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  • Hosted by:Chinese Academy of Sciences
    Sponsored by:Institute of Botany, Chinese Academy of Sciences, Botanical Society of China
    Co-hosted by:Key Laboratory of Soybean Molecular Design Breeding, Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences
    Institute of Biotechnology and Germplasm Resources, Yunnan AgriculturalAcademy
    Fujian Agriculture and Forestry University
    Hunan Provincial Key Laboratory of Phytohormones and Growth Development, Hunan Agricultural University
    State Key Laboratory of Crops Biology, Shandong Agricultural University

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Methods for TurboID-based Proximal Labeling in Plants
Jiayi Kuang, Hongqing Li, Wenjin Shen, Caiji Gao
Chinese Bulletin of Botany    2021, 56 (5): 584-593.   DOI: 10.11983/CBB21104
Accepted: 11 August 2021

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Proximity labeling (PL), a recently developed technique to detect protein-protein interactions and subcellular structural proteomes in living cells, has been successfully applied in various animal and plant systems. Proximity labeling is conducted by fusing an engineered enzyme with catalytic activity to a protein of interest (bait protein). With the catalysis of the enzyme, small molecular substrates such as biotin are covalently linked to endogenous proximal proteins, which can be further enriched and analyzed to identify the interactome of the bait protein. TurboID, a biotin ligase produced by directed evolution, has the advantages of non-toxicity and high catalytic efficiency. Using TurboID-based proximity labeling to analyze proximal proteins of bait proteins, we can study transient or weak protein interactions, which helps to understand the complex biological processes occurring inside cells. Here, we describe methods and related tips for TurboID-based proximal labeling in Arabidopsis thaliana, and hope to provide a reference for studying plant protein-protein interactions.

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Advances in Biological Functions of Aux/IAA Gene Family in Plants
Yanyan Li, Yanhua Qi
Chinese Bulletin of Botany    2022, 57 (1): 30-41.   DOI: 10.11983/CBB21168
Accepted: 28 December 2021

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Auxin is one of the most important plant hormones and plays a key role in regulating plant growth and development. In plants, early auxin responsive gene families, such as Aux/IAA (Auxin/Indole acetic acid repressors), GH3 (Gretchen Hagen3) and SAUR (Small Auxin up RNA), are rapidly induced and up-regulated by auxin treatment. Aux/IAA gene family is generally composed of four conserved domains. Domain I inhibits the expression of downstream genes in the auxin signaling pathway, and domain II is mainly regulated by Transport Inhibitor Response 1 (TIR1) in auxin signal transduction, thus affecting the stability of Aux/IAA. Domain III/IV regulates auxin signaling by interacting with Auxin Response Factor (ARF). Aux/IAA gene family has been reported to play an important role in organ development, root formation, stem elongation and leaf expansion in dicotyledonous Arabidopsis thaliana while in monocotyledonous rice (Oryza sativa) and wheat (Triticum aestivum), Aux/IAA mainly affects root development and plant architecture. However, the functions of most Aux/IAA genes remain unclear and need to further study. In this article, we reviewed the structure and function of Aux/IAA protein, and the auxin signal transduction pathway in Arabidopsis, cereal crops and other plants to provide clues for fully revealing the biofunction of the Aux/IAA gene family.

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The Gold Will Glitter Wherever it is: Convergent Selection in Maize and Rice
Hong Yu, Jiayang Li
Chinese Bulletin of Botany    2022, 57 (2): 153-156.   DOI: 10.11983/CBB22054
Accepted: 24 March 2022

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Domestication of wild plants was crucial for human settlement and the development of civilization, which arose independently in many different geographic areas on different wild species. However, these crops underwent variant domestication process displaying the ‘domestication syndrome’ with a common suite of traits. The systematical analysis of convergent selection at genome level may provide important information and genetic resources for crop breeding. Recently, a team led by Xiaohong Yang and Jiansheng Li from Chinese Agricultural University and Jianbing Yan from Huazhong Agricultural University reported the genetic basis of convergent selection between maize and rice at both single gene and whole genome levels. Particularly, they found the maize KRN2 and rice OsKRN2 genes experienced convergent selection and regulated grain number and yield in a similar pathway. Moreover, they identified a large number of orthologous gene pairs that underwent convergent selection during maize and rice evolution, which were enriched in certain pathways including starch metabolism, sugar and coenzyme synthesis. This significant work not only cloned KRN2/OsKRN2 orthologous gene pairs with great value in maize and rice breeding, but also revealed the convergent selection between maize and rice at the genome level, providing critical foundations for studying the molecular basis of domestication syndrome and their applications in breeding practices.

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Progress in Flowering Regulation Mechanisms of FLC
Changsheng Zhang, Tao Wei, Yuping Zhou, Tian Fan, Tianxiao Lü, Chang'en Tian
Chinese Bulletin of Botany    2021, 56 (6): 651-663.   DOI: 10.11983/CBB21103
Accepted: 10 September 2021

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FLOWERING LOCUS C (FLC) is an important repressor of plant flowering. It mainly represses flowering by binding to the promoters of two downstream key floral promoting genes, FLOWERING LOCUS T (FT) and SUPPRESSOR OF OVEREXPRESSION OF CO 1 (SOC1). It also regulates flowering time by binding to other regulatory genes. While the detailed molecular mechanisms of FLC in the regulation of flowering still need to be deeply investigated. This review summarizes the new research progresses of FLC in recent years in the 8 genetic pathways of flowering regulation, and give an outlook for the future study in this field.

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Effects of 60Co-γ Ray Radiation on Morphology and Volatile Components of Origanum vulgare
Yihao Yan, Di Wang, Jingyi Li, Wenying Zhang, Yuanpeng Hao, Fei Xia, Hui Li, Hongtong Bai, Lei Shi
Chinese Bulletin of Botany    2021, 56 (5): 533-543.   DOI: 10.11983/CBB21012
Accepted: 21 April 2021

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Carvacrol and thymol, the main components of essential oil in oregano (Origanum vulgare), have strong antibacterial activity, and have big potential to replace antibiotics. In order to acquire valuable genetic variation in oregano,60Co-γ ray radiation was utilized in breeding of oregano. The volatile components were analyzed by SPME-GC-MS, also principal component analysis and hierarchical clustering composition were performed. The effects of60Co-γ ray radiation on morphological characteristics were studied, including glandular hair density, glandular hair size and volatile components of M1 generation of oregano. The results showed that the LD50 of oregano seeds was 16.39 Gy; The plant height, stem diameter, branching number, leaf length and width of irradiated M1 generation were all changed, and several morphological mutants were selected; At the same time, the density and size of glandular hair changed in a variety of ways, and 25 mutants with significantly increase in density and size of glandular hair were selected from 163 surviving plants;60Co-γ ray treatment had little effect on volatile components, but significant effect on the content of volatile components, and 5 chemotypes in oregano were induced, including carvacrol-type, thymol-type, γ-terpinene-type, β-caryophyllene- type, and germacrene-type. Six mutants with high carvacrol and thymol were obtained. These results confirmed that60Co-γ ray radiation could be useful as an effective mutagenic method for oregano breeding, and the suitable radiation dose was 20 Gy. This study provided basic data and new way for the breeding and selection of new germplasm resources in oregano.

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Cited: CSCD(1)
Achievements and Advances in the Plant Sciences in China in 2021
Fan Chen, Hongya Gu, Xiaoquan Qi, Qian Qian, Jianru Zuo, Shuhua Yang, Zhiduan Chen, Lei Wang, Rongcheng Lin, Liwen Jiang, Xiaojing Wang, Langtao Xiao, Yongfei Bai, Kang Chong, Tai Wang
Chinese Bulletin of Botany    2022, 57 (2): 139-152.   DOI: 10.11983/CBB22090
Accepted: 26 April 2022

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In 2021, the numbers of original research articles published by Chinese plant scientists in international multidisciplinary journals and mainstream plant science journals increased significantly compared with that in 2020, and important advances have been made in the fields of male and female cell recognition and fertilization, stem cell fate determination, mycorrhizal symbiosis, photosynthetic membrane protein complex, nitrogen and phosphorus nutrient utilization, innate immunity, crop de novo domestication and genome design. Among them, ‘rapid de novo domestication of allotetraploid wild rice’ was selected as one of the ‘Top Ten Advances in Life Sciences in China’ in 2021. Here we summarize the achievements of plant science research in China in 2021, and briefly introduce 30 representative important research advances, so as to help readers understand the developmental trend of plant sciences in China, and evaluate how to better connect plant scientific research with major national needs.

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Research Progress of Melatonin in Plant Stress Resistance
Deshuai Liu, Lei Yao, Weirong Xu, Mei Feng, Wenkong Yao
Chinese Bulletin of Botany    2022, 57 (1): 111-126.   DOI: 10.11983/CBB21146
Accepted: 17 December 2021

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Melatonin (N-acetyl-5-methoxytryptamine) is a small molecule indoleamine that is essential for life. Melatonin is widely present in animals and plants, and plays a vital role in the growth and development. With the study of plant melatonin, its synthesis pathway and roles in plants have become clear. Studies have shown that melatonin has the regulatory effects in improving plant resistance to abiotic and biotic stresses. This review summarizes the research progress of melatonin in plant abiotic and biotic stresses in recent years; it provides reference for in-depth analysis of influence of melatonin on the regulation mechanism of plant resistance to stresses.

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Research Progress on Apoplast Barriers of Plant Roots
Xin Liu, Pei Wang, Qingping Zhou
Chinese Bulletin of Botany    2021, 56 (6): 761-773.   DOI: 10.11983/CBB21054
Accepted: 18 June 2021

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Root is the most important organ of plants to absorb water and mineral nutrients for maintaining life activities. The architecture and ultrastructure of roots are species specific with different effects on the water and mineral nutrients absorption. Among them, suberin lamellae and Casparian strip in the endodermis and exodermis are two important apoplast barriers, which block water and ions transport nondirectionally and play important roles in the plant growth and response to stress. However, the structure, chemical composition, physiological functions, biosynthesis and regulation of root apoplast barriers are only studied in Arabidopsis thaliana. In recent years, studies of root apoplast barriers in barley (Hordeum vulgare), rice (Oryza sativa) and some forages have just been reported. In this review, the research progress of root apoplast barriers in A. thaliana, barley, rice and some forages in recent years are compared, and the future research direction is put forward in order to provide a theoretical basis for further exploring the roles of apoplastic barriers of gramineous crops and forages in growth, development and stress adaptation, and provide new ideas for crop and forage breeding.

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Chromatin Accessibility and the Gene Expression Regulation in Plants
Zhanjie Li, Yuan Qin
Chinese Bulletin of Botany    2021, 56 (6): 664-675.   DOI: 10.11983/CBB21115
Accepted: 12 October 2021

Abstract528)   HTML187)    PDF (1375KB)(329)       Save

The topological organization of nucleosomes across the genome is non-uniform. While densely arranged within constitutive heterochromatin, histones are depleted at regulatory loci. Chromatin accessibility is the degree to which nuclear macromolecules are able to physically contact with regulatory DNA. Following the development of next-generation sequencing technology, a variety of quantitative methods, including DNase-seq, ATAC-seq, MNase-seq and NOMe-seq, have been developed to measure genome-wide chromatin accessibility easily and efficiently. In this review, we first introduced the technical principles of the four principal methods for measuring chromatin accessibility. And then we summarized the critical biophysical determinants of chromatin accessibility, including nucleosome occupancy, histone modification and TFs combination. Finally, we described recent advances of chromatin regulation during development and stress responses in plants. Our goal is to provide a reference for researches about genome-wide chromatin accessibility mapping, identification of cis-regulatory elements, and the dissection of epigenetic and genetic regulatory networks.

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Regulatory Mechanisms of the Plant CBL-CIPK Signaling System in Response to Abiotic Stress
Lingling Xie, Jinlong Wang, Guoqiang Wu
Chinese Bulletin of Botany    2021, 56 (5): 614-626.   DOI: 10.11983/CBB21024
Accepted: 21 April 2021

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Calcineurin B-like proteins (CBLs) and their CBL-interacting protein kinases (CIPKs) are important regulatory network in response to abiotic stresses. The CBL-CIPK system senses and decodes Ca2+-signals through phosphorylation to regulate plant response to abiotic stresses. In this review, the basic structures of CBLs and CIPKs, and their phosphorylation on different substrates, as well as regulatory mechanisms of plants in response to abiotic stresses were summarized. We also put forward a perspective on the future research directions of CBLs and CIPKs, as well as their potential applications in genetic improvement of crops for stress tolerance.

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Map-based Cloning and Natural Variation Analysis of the PAL3 Gene Controlling Panicle Length in Rice
Jiangyuan Shang, Yan Chun, Xueyong Li
Chinese Bulletin of Botany    2021, 56 (5): 520-532.   DOI: 10.11983/CBB21119
Accepted: 16 September 2021

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Panicle architecture is one of the key factors determining the rice (Oryza sativa) grain yield. In this study, four short panicle mutants were identified from an EMS (ethyl methane sulfonate) mutant library of a japonica vareity Shengdao 808 (SD808). The panicle length, primary branch number, secondary branch number and grain number per panicle of these mutants were decreased in various degrees. Map-based cloning showed that these mutants were controlled by the gene PAL3 (PANICLE LENGTH 3) which encodes a peptide transporter with 12 transmembrane domains. The point mutations of pal3-1 and pal3-2 resulted in non-synonymous mutations of amino acids in the conserved region; the point mutation of pal3-3 resulted in the mis-splicing of the first exon and intron; and the point mutation of pal3-4 resulted in the premature termination of translation and thus the deletion of the 12th transmembrane domain. Through haplotype analysis, nine haplotypes of PAL3 were identified, including three major haplotypes (Hap1-Hap3). Hap1 is dominated by japonica accessions, Hap2 contains both indica and japonica accessions, while Hap3 is dominated by indica accessions. Hap1 originated from O. rufipogon, while Hap2 and Hap3 may originate from O. nivara. Statistical analysis showed that the panicle length of Hap3 was significantly higher than that of Hap1 and Hap2, indicating that Hap3 may have the potential to improve the panicle length. In conclusion, this study revealed the important role of the peptide transporter in regulating rice panicle architecture and thus provides a new theoretical basis for rice panicle architecture improvement.

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Conquering the Summit: A New Era Towards Hybrid Seed Potato Breeding
Cao Xu
Chinese Bulletin of Botany    2021, 56 (5): 516-519.   DOI: 10.11983/CBB21151
Accepted: 17 September 2021

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Modern cultivated potato (Solanum tuberosum) is a clonally propagated autotetraploid, with highly heterozygous genome, complex genetic background, and severe inbreeding depression, making it difficult to combine eminent traits and resulting in a long breeding cycle and the dilemma of ‘genetic stagnation’ of potato hybrid breeding. Moreover, clonal propagation leads to low reproduction coefficient, high cost of storage and transportation, whereas tubers are easy to carry viruses and pests, which have hindered the development of potato industry for a long time. Recently a team led by Sanwen Huang in Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, successfully use genome design to develop the pure and fertile potato lines and thereby the hybrid F1, reinvent potato from a clonally propagated tetraploid into a seed-propagated diploid. This work is a milestone in potato breeding, that starts a new era of genome design and rapid potato breeding.

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Cited: CSCD(1)
Novel Links in the Plant Target of Rapamycin Signaling Networks
Yanyan Meng, Nan Zhang, Yan Xiong
Chinese Bulletin of Botany    2022, 57 (1): 1-11.   DOI: 10.11983/CBB21183
Accepted: 10 January 2022

Abstract493)   HTML45)    PDF (453KB)(680)       Save

Target of rapamycin (TOR) is a highly conserved serine/threonine-protein kinase among all eukaryotes. As a central regulator, TOR integrates nutrient, energy, growth factor and environmental signals to control cell proliferation, growth, and metabolism. With the establishment of TOR research systems in plants, tremendous progress has been made in, both conserved and unique functions of plant TOR have been uncovered in recent years. Here, we reviewed TOR complexes and the mechanisms of plant TOR integrating sugar, nitrogen, phosphate, sulfur, phytohormone and stress signals to orchestrate transcription, translation, metabolism, autophagy, and stress responses. We also highlight a few fundamental questions that will be of great interest to be resolved for fully revealing biofunctions of plant TOR.

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Advance in Gene-editing Technology Based on CRISPR/Cas9 and Its Application in Plants
无 无无 xifeng Chen
Chinese Bulletin of Botany    2022, 57 (4): 0-0.   DOI: 10.11983/CBB22020
Accepted: 24 April 2022

Abstract467)      PDF (3012KB)(528)       Save
CRISPR/Cas9 technology is a gene editing strategy using Cas9 nuclease guided by RNA to target an interest gene in genome. Recently, a large number of new-type gene editors based on the CRISPR/Cas9 have been updated rapidly and become more precisely and efficiently in gene editing, which has a great application prospect in crop molecular-designing breeding. This paper summarizes the technical principles, editing effects and applications of the CRISPR/cas9 and its related gene editors, and also discusses the aspects of dilemmas, countermeasures and prospects, intending to provide reference for scientific researchers in the related fields.
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Research Progress of Hyperhydricity Mechanism in Plant Seedling Growth
Qingping Zhao, Yuping liang, Fangyuan Zhou, Xiang Zhao
Chinese Bulletin of Botany    2022, 57 (1): 90-97.   DOI: 10.11983/CBB21126
Accepted: 12 October 2021

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Hyperhydricity (HH) or vitrification is a physiological disorder during plant growth, which shows a typical translucent phenotype like water-soaked that often occurs in the plant tissue culture. Vitrification is one of the three main influencing factors that limit the tissue culture technology and the commercialized production of seedlings during the plant tissue culture, but its molecular mechanism is still unclear by far. Using plantlets from the tissue culture as a research object often lead to an interference of man-made factors, which can be avoided by non-tissue culture material to reveal the molecular mechanism of vitrification in nature. This review summarized the recent progress of mechanism for inducing HH in non-tissue culture plants, including the abnormal deposit of suberin, the reduction of cuticular wax, the lipid peroxidation in cellular membranes and the disrupted transmembrane transport of ion or water, to provide new clues and thinking for hyperhydricity in plant seedling growth.

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Recognition, Immune Activation and Signal Regulation of Plant NLR Immune Receptor
Lei Qin, Zhihong Peng, Shitou Xia
Chinese Bulletin of Botany    2022, 57 (1): 12-23.   DOI: 10.11983/CBB21159
Accepted: 07 February 2022

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A large area of membrane surface and intracellular immune receptors have been evolved in higher plants to sense various pathogen signals and prevent pathogen invasion. Among them, pattern recognition receptors on the cell surface activate basic immune response after sensing pattern molecules, while nucleotide-bounding leucine-rich repeat proteins (NLRs) activate specific immune response by sensing effector proteins secreted by pathogenic microorganisms, resulting in hypersensitivity and cell death. In this review, the latest research progress of plant immunity is mainly reviewed from the aspects of NLRs on the recognition of effector proteins, plant immune activation and downstream signal regulation.

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Research Progress of the Physiological and Molecular Mechanisms of Cadmium Accumulation in Rice
Luyao Wang, Jian Chen, Shouqing Zhao, Huili Yan, Wenxiu Xu, Ruoxi Liu, Mi Ma, Yijun Yu, Zhenyan He
Chinese Bulletin of Botany    2022, 57 (2): 236-249.   DOI: 10.11983/CBB21222
Accepted: 20 January 2022

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The wide occurrence of Cadmium (Cd)-contaminated rice in China poses significant public health risk. Breeding rice varieties with low-Cd accumulation is an effective strategy to reduce Cd accumulation in rice grain. It is necessary to understand the characteristics of Cd accumulation in rice, its physiological process and related functional genes. Here, we review the advances in physiological and molecular mechanisms of Cd uptake in roots, loading and translocation in xylem, distribution in nodes, redistribution in leaves, and accumulation in grains, which will provide theoretical reference for breeding and safe production of low-Cd rice.

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A Ca2+-ROS Signaling Axis in Rice Provides Clues to Rice-pathogen Coevolution and Crop Improvements
Jian-Min Zhou
Chinese Bulletin of Botany    2021, 56 (5): 513-515.   DOI: 10.11983/CBB21160
Accepted: 30 September 2021

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Chinese scientists have made multiple breakthroughs in recent years in rice disease resistance studies, particularly in the areas of durable resistance and resistance-yield coordination. Most recently, a team led by Zuhua He and Weibing Yang at the CAS Center for Excellence in Molecular Plant Sciences made another major advance in our understanding of disease resistance and host-pathogen co-evolution in rice. They showed that the calcium sensor protein ROD1 directly enhances the activity of catalase protein CatB to remove reactive oxygen species during immune responses, preventing excessive immune responses and ensuring optimum rice plant growth. Remarkably, they also illustrated that a functionally attenuated variant of ROD1 is enriched in wild and domesticated Indica rice from tropical and subtropical regions and that this variant allows elevated resistance against sheath blight disease. Importantly, this variant is likely useful for breeding as it does not compromise rice growth and yield. Interestingly, they further demonstrated that an effector protein from the rice blast fungus structurally and functionally mimics ROD1 to suppress rice immunity. Thus, this study uncovers an immune regulatory axis defined by ROD1 and CatB that is at the center of rice-pathogen co-evolution.

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Cited: CSCD(1)
Establishment and Application of a Rapid Genetic Transformation Method for Peanut
Qiong Zhai, Rongqin Chen, Xiaohua Liang, Chuchun Zeng, Bo Hu, Ling Li, Xiaoyun Li
Chinese Bulletin of Botany    2022, 57 (3): 327-339.   DOI: 10.11983/CBB21192
Accepted: 18 March 2022

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Plant transformation is an important tool for genetic engineering. The key technology of genetic transformation is to introduce foreign genes into plants genomes quickly and efficiently and reduce the time to obtain transgenic offspring. Peanut (Arachis hypogaea) is an important oil and cash crop in China. The genetic transformation system is still not well established in peanuts. It seriously restricts the function research of peanut genes and the molecular breeding progress. Here, we established a rapid and stable genetic transformation system in peanuts. The Agrobacterium tumefaciens was injected into the second stem of peanut to produce transgenic plants. Then positive transgenic peanut was transplanted and tamped backfill to cover the injection point. Those seeds above the injection point were picked for further screening and analyzing. The results showed that over 40% of transgenic plants were obtained and displayed chimeric in T0 generation. The T0 seeds were harvested about 5 months after rapid-transformation. About 9% of T1 peanuts were hybrids rather than chimeras. To solve the problem of few seed in some transgenic plants, the rapid-transformation system was combined with traditional tissue culture. This rapid-transformation system has potential value in garlic (Allium sativum), potato (Solanum tuberosum), and freesia (Freesia refracta). Altogether, this study establishes a rapid and stable genetic transformation system for peanuts, which sheds light on other plants’ genetic transformation.

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Analysis on the Evolution and Transcription Activation Activity of ABI4 in Brassicaceae
Wenjing Zhang, Xiaomeng Yang, Kan Gao, Xinyi Wei, Xuetong Shi, Ruixuan Wang, Fengxia Wu, Juqing Kang
Chinese Bulletin of Botany    2021, 56 (6): 676-686.   DOI: 10.11983/CBB21036
Accepted: 11 August 2021

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ABI4 is an important component of the ABA signal transduction pathway. It not only acts as a key player in ABA and GA antagonism, but also plays important roles in various aspects of ABA crosstalk with other signal chemicals. Genes regulated by ABI4 are involved in diverse processes. In this study, we got 27 homologs from 19 species in the family Brassicaceae using AthABI4 as a query, and explored their evolutionary history based on sequence polymorphism analysis, phylogenetic reconstruction, genomic synteny assay, and transcription activation activity comparison. The result revealed that ABI4 are highly conserved and might be not work as transcriptional activators independently in the Brassicaceae, which implies that the irreplaceable roles of ABI4 in plants and further research was needed to explore the molecular mechanism of its biological function.

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The Role of Alternative Splicing in Floral Transition
Fangfang Cai, Changsheng Shao, Yuqiang Sun
Chinese Bulletin of Botany    2022, 57 (1): 69-79.   DOI: 10.11983/CBB21085
Accepted: 12 October 2021

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Precise regulation of floral transition, which ensures plants bloom in the suitable environment, is essential for the successful reproduction of plants. The flowering time genes are regulated by a variety of molecular mechanisms, including transcriptional, post-transcriptional and post-translational regulations. Alternative splicing is a universal molecular process at the post-transcriptional level that can generate multiple transcripts from a single gene, thereby enriching the diversity of the transcriptome and proteome. The accumulating evidence indicates that alternative splicing plays an important role in the floral transition. According to developmental and environmental cues, alternative splicing can regulate the levels of functional transcripts and/or proteins of flowering time genes by affecting the stability of mRNA and/or the function of protein isoforms. Therefore, revealing the roles of alternative splicing will further improve our understanding of the functions of flowering time genes and the whole regulatory network of floral transition. In this review, we introduce the research progress of alternative splicing in floral transition, and summarize from the various regulatory pathways, thereby providing a reference for further research on the regulatory mechanisms of alternative splicing and floral transition in plants.

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An Efficient Protoplast Transient Expression System in Camellia sinensis var. sinensis cv. ‘Tieguanyin’
Yuqin Zhang, Jiacheng Wu, Meng He, Renyi Liu, Xiaoyue Zhu
Chinese Bulletin of Botany    2022, 57 (3): 340-349.   DOI: 10.11983/CBB21206
Accepted: 07 February 2022

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Recent advances in the genomic sequencing of tea plants have laid the foundation for tea research at the molecular and gene levels. However, the transgenic technologies are immature and the life cycles are long for tea trees, it is still difficult to conduct functional analyses of tea genes. This study used young leaves of Camellia sinensis var. sinensis cv. ‘Tieguanyin’, established a useful formula by testing multiple concentration combinations of cellulase, pectinase, macerozyme and mannitol. By evaluating the quantity, viability and debris of resulted protoplast, we successfully established a highly efficient mesophyll protoplast isolation and PEG-mediated transient expression system in Tieguanyin seedling leaves, with a transformation rate reaching 56.25%. Using this system, the subcellular localization of two pivotal enzymes in the theanine metabolism pathway (the theanine synthetase (TSI) and the glutamine synthetase (GSII-1.1)) were explored. Results show that, these two enzymes are both localized in the cytosol of the Tieguanyin protoplasts. Together, the establishment of this tea mesophyll protoplast extraction and transient expression system would lay a technological foundation for studying the function of tea genome.

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Managing Both Internal and Foreign Affairs—A PHR-centered Gene Network Regulates Plant-mycorrhizal Symbiosis
Dong Liu
Chinese Bulletin of Botany    2021, 56 (6): 647-650.   DOI: 10.11983/CBB21177
Accepted: 26 October 2021

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Phosphorus is a macronutrient essential for plant growth and development, however, phosphate (Pi), the major form of phosphorus absorbed by plants, is quite limiting in soil. To cope with this nutritional stress, plants have evolved an array of adaptive responses, which are largely regulated by changing gene expression in response to Pi deficiency. The transcription factor, PHR1 plays a key role in regulating plant transcriptional response to Pi deficiency. Besides, most land plants can form symbiosis with arbuscular mycorrhizal (AM) fungi, through which plants can obtain Pi from soil more effectively. Recently, the research group of Ertao Wang of Center for Excellence in Molecular Plant Science, Chinese Academy of Sciences, reported that a PHR-centered gene regulatory network plays an essential role in promoting plant-AM symbiosis. Therefore, PHR not only functions in maintaining plant Pi homeostasis, but also in communicating with beneficial microorganisms in the environments, which provides another route for plants to obtain Pi from soil.

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Cited: CSCD(1)
Advances in Three-dimensional Characteristics of Photosynthesis in Plants
Qingqing Zou, Hanyu Wu, Donghuan Liu, Chuangdao Jiang
Chinese Bulletin of Botany    2022, 57 (2): 250-258.   DOI: 10.11983/CBB21213
Accepted: 25 February 2022

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Photosynthesis is the most important chemical reaction on earth. Though photosynthesis in plant has been extensively studied, little attention has been paid to the photosynthetic function of plant leaves and its’ regulation in three-dimensional level. There are obvious three-dimensional characteristics in leaf structure, photosynthetic components, light transmission, and light absorption in leaves, which may greatly affect the CO2 transport inside leaves, electron transport and carbon assimilation in various mesophyll cells. Unavoidably, the photosynthetic function and its’ regulation in leaves may show complex three-dimensional characteristics. Consequently, the analysis of leaf photosynthetic characteristics from the three-dimensional perspective will help us understand the mechanism of photosynthesis and provide a theoretical support to improve photosynthetic efficiency in plants.

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Establishment of Tissue Culture System for Axillary Bud Regeneration of Primula × pubescens
Mengyue Li, Liu Liu, Yan Liu, Xiaoman Zhang
Chinese Bulletin of Botany    2021, 56 (6): 732-739.   DOI: 10.11983/CBB21076
Accepted: 30 August 2021

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The effects of different concentrations of hormone on Primula × pubescens tissue culture were studied through callus induction and differentiation, adventitious bud proliferation culture and rooting. The optimal medium at different stages was selected to optimize the regeneration tissue culture system of Primula × pubescens. Our results showed that the best medium for callus induction and differentiation was MS+0.2 mg∙L-1 NAA+1.0 mg∙L-1 6-BA, with 84% induction rate and 67% budding rate; the optimal medium for adventitious bud proliferation was MS+0.5 mg∙L-1 NAA+0.2 mg∙L-1 6-BA, and the proliferation rate was 67%; MS+0.2 mg∙L-1 NAA medium was the best for rooting and elongation of tissue culture seedlings. The average number of roots per plant was 9, and the rooting rate was 70%. This study established a highly effect tissue culture and regeneration system of Primula × pubescens, which laid a foundation for genetic research and germplasm innovation of other Primula plants.

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QTL Mapping and Candidate Gene Analysis of Vitamin E in Rice Grain
Hanfei Ye, Wenjing Yin, Yian Guan, Kairu Yang, Qianyu Chen, Shuying Yu, Xudong Zhu, Dedong Xin, Wei Zhang, Yuexing Wang, Yuchun Rao
Chinese Bulletin of Botany    2022, 57 (2): 157-170.   DOI: 10.11983/CBB21216
Accepted: 20 January 2022

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Vitamin E (VE) is an important index of rice nutritional quality. Rice (Oryza sativa) is the most widely planted crop in China. To increase rice VE content is a convenient and effective way to improve its nutrient composition. In this study, 120 recombinant inbred line (RIL) populations were constructed with indica rice HZ as male parent and japonica rice Nekken2 as female parent. The content of vitamin E isomer in the population was determined by high performance liquid chromatography (HPLC). Quantitative trait loci (QTL) mapping was carried out based on the constructed high-density molecular genetic map. After pedigree analysis, 122 VE related QTLs were identified, which were distributed on 12 chromosomes. Some loci have high LOD scores, such as qT3α/to2-1 with 10.32 LOD score, qT3α2-1 of 9.91 LOD score. There are multiple major QTLs associated with the content of different conformations, and some VE biosynthesis genes are within the intervals, e.g., OsGGR1, OsGGR2, OsTC and OsγTMT. The expression of these genes were analyzed by qRT-PCR. The results showed that their expression in HZ was significantly higher than that in Nekken2. It is speculated that the high expression of these genes is the reason of why tocopherol and tocotrienol in HZ are higher than in Nekken2. The number of QTLs and the scores of LOD are large, which provides a molecular basis for further screening and cultivating rice varieties with high VE content also conductive to clarify the molecular regulation mechanism of rice VE biosynthesis through QTLs.

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Analysis on the Data of Airborne Pollen in Beijing-Tianjin-Hebei Region
Dejingyu Zhao, Caihua Ye, Yufei Wang, Yifeng Yao
Chinese Bulletin of Botany    2021, 56 (6): 751-760.   DOI: 10.11983/CBB21061
Accepted: 30 August 2021

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As one of the components of urban air pollutants, pollen seriously affects our living environment and health, which is highly concerned by government and scientific community. Here, based on the airborne pollen data of past 60 years in Beijing-Tianjin-Hebei region, we summarize the major pollen types and their seasonal distribution characteristics in this region, and the data shows that the annual variation of airborne pollen concentration basically follows the ‘bimodal' pattern, viz. arboreal pollen such as Cupressaceae, Salicaceae and Betulaceae dominate the peak in spring, while herbaceous pollen like Artemisia, Humulus/Cannabis sativa, Chenopodiaceae/Amaranthaceae dominate the peak in summer and autumn. Then we discuss the dominant meteorological factors related to pollen concentration and the characteristics of pollinosis. Furthermore, we point out that human activities (e.g., land reconstruction and roadside tree planting) may have an impact on the changes of airborne pollen composition in Beijing over these years. Finally, we emphasize the important role of future long-term airborne pollen monitoring in atmospheric environment assessment, pollinosis prevention, and urban greening construction.

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In Vitro Rapid Propagation of Nelumbo nucifera
Yaqian Xiong, Xianbao Deng, Huihui Zhang, Dong Yang, Heng Sun, Juan Liu, Mei Yang
Chinese Bulletin of Botany    2021, 56 (5): 605-613.   DOI: 10.11983/CBB21020
Accepted: 27 May 2021

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Using fresh embryos isolated from developing lotus seeds of 18 days after pollination as explants, a rapid propagation system of lotus was established through primary culture, subculture plantlets hardening and transplantation. Results showed that the primary culture medium of MS+0.5 mg·L-1 6-BA+0.5 mg·L-1 NAA+30 g·L-1 sucrose+0.5 g·L-1 activated carbon+0.8 g·L-1agar was the optimum for the explant growth. The explant induction rate was up to 85% after cultured for 60 days, Qiuhongyang had the highest amount of stem nodes. In the subculture, the optimal concentration of sucrose in the medium was 80 g·L-1. When dividing aseptic seedling for subculture, cutting with two stem nodes showed the highest multiplication coefficient. The multiplication coefficient of different varieties was ranged from 4.0 to 6.7, and Qiuhongyang was the highest (6.7). Aseptic seedlings were suggested to be subcultured every 50 days, with high multiplication rates for up to 6 rounds of subcultures. The rooted plantlets were transplanted to pots containing medium of peat:pond soil=1:2 (v/v) during May to July, and the survival rate of transplanted plants was higher than 83.9%. Based on these results, using this in vitro propagation system, it is estimated that 1 465 seedlings be developed from a single lotus seed within one year. This study has established a rapid in vitro propagation system for lotus, which provides the basis for the factory scale production of lotus plantlets.

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Identification and Mapping of a Rice Male Sterility Mutant ms102
Xia Wang, Wei Yan, Zhiqin Zhou, Zhenyi Chang, Minting Zheng, Xiaoyan Tang, Jianxin Wu
Chinese Bulletin of Botany    2022, 57 (1): 42-55.   DOI: 10.11983/CBB21158
Accepted: 17 November 2021

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The recessive nuclear male sterility line is the base of the third-generation hybrid rice breeding technology. To explore new male sterility lines suitable for the breeding technology, we screened an ethyl methylsulfonate (EMS) mutant library derived from the elite xian variety Huanghuazhan (HHZ) and identified male sterility mutant 102 (ms102). The ms102 showed a normal vegetative development, but exhibited defective anther dehiscence and pollen abortion. Cytological analyses revealed that the degradation of the tapetum was abnormal in ms102, resulting in the abortion of microspores. Genetic analyses showed that the male sterile phenotype of ms102 was resulted from the mutation of a known male sterility gene Defective Pollen Wall 2 (DPW2), which encodes an acyl transferase. Thus, this study identified a recessive nuclear male sterility mutant, and further confirmed the function of DPW2 in rice pollen development.

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Advance in Molecular Mechanism of MBF1 Regulating Plant Heat Response and Development
Yi Qin, Yanshuang Liu, Liuliu Qiu, Min Zhou, Xiaoshan Du, Shaojun Dai, Meihong Sun
Chinese Bulletin of Botany    2022, 57 (1): 56-68.   DOI: 10.11983/CBB21220
Accepted: 19 January 2022

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MBF1 (multiprotein bridging factor 1) is a transcriptional co-activator, which is highly conserved in evolution. It exists in all eukaryotes and can activate gene transcription by connecting components of the basal transcription machinery and transcription factors. Plant MBF1 plays important roles in a variety of biological processes, including controlling plant growth and development, and adversity adaptation. This review demonstrates the advance in the molecular structure and regulation mechanism of plant MBF1, and highlights the molecular mechanism of AtMBF1c in the regulation of plant heat stress response.

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Cloning and Functional Analysis of Rice Yellow Green Leaf Regulatory Gene YGL18
Kairu Yang, Qiwei Jia, Jiayi Jin, Hanfei Ye, Sheng Wang, Qianyu Chen, Yian Guan, Chenyang Pan, Dedong Xin, Yuan Fang, Yuexing Wang, Yuchun Rao
Chinese Bulletin of Botany    2022, 57 (3): 276-287.   DOI: 10.11983/CBB22018
Accepted: 18 March 2022

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Leaf color mutants are often accompanied by changes in chlorophyll content and abnormal chloroplast structure, and serve as essential materials for studying the functions of chloroplast development and photosynthesis-related genes. In this study, we obtained a yellow-green leaf mutant named yellow-green leaf 18 (ygl18) from Oryza sativa subsp. indica cv. ‘HZ’ with ethyl methanesulfonate (EMS). Compared with the wild type, the leaves of ygl18 turned yellow at three-leaf stage and the degree of yellowing increased as it grew, accompanied by decreasing photosynthetic rate and chlorophyll content. The seed-setting rate, 1 000-grain weight, and effective panicle number were significantly lower than those of the wild type. We observed disordered chloroplast structure, loose stromal lamellas, and stalled development in the mutant using transmission electron microscopy. Genetic analysis indicated that the mutant feature (or phenotype) of ygl18 is controlled by a pair of recessive nuclear alleles, which were located in a 115.2 kb region between markers InDel2 and InDel3 on the long arm of chromosome 3. We found mutations in the 5′UTR of LOC_Os03g48040 encoding FdC2 (ferredoxin C2). The gene’s function on controlling the mutant phenotype was verified using CRISPR transgenic experiments. Our results revealed a genetic basis for leaf color regulatory network and provide new clues for breeding photosynthetically efficient rice varieties in the future.

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Alternative splicing of plant genes: Full of change, Sail with wind
Xuefeng Hao, Zhangjing Wu, Tian Ma, Zhuping Jin, Yanxi Pei
Chinese Bulletin of Botany    DOI: 10.11983/CBB22041
Accepted: 10 May 2022

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Rice OsWRKY42 is a Novel Element in Xa21-mediated Resistance Pathway Against Bacterial Leaf Blight
Tianxingzi Wang, Zheng Zhu, Yue Chen, Yuqing Liu, Gaowei Yan, Shan Xu, Tong Zhang, Jinjiao Ma, Shijuan Dou, Liyun Li, Guozhen Liu
Chinese Bulletin of Botany    2021, 56 (6): 687-698.   DOI: 10.11983/CBB21025
Accepted: 11 August 2021

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Xa21 was the earliest resistant gene cloned in rice, which has broad-spectrum resistance to Xanthomonas oryzae pv. oryzae (Xoo). Our previous study demonstrated that, OsWRKY42 might play a role in Xa21-mediated resistance pathway. In this study, OsWRKY42-RNA interference (OsWRKY42-RNAi) transgenic rice were generated under the background of Xa21, Western blot (WB) verified transgenic plants were used to inoculate Xoo, it was found that the lesion length was longer than that resistance control 4021 (carrying Xa21 gene), indicating that the down-regulation of OsWRKY42 protein abundance impaired Xa21-mediated resistance. In OsWRKY42-RNAi transgenic rice, it was also found by WB analysis that the abundance of OsPR6, OsPR15 and OsPR16 protein were decreased, while the abundance of OsPR1A, OsPR1B, OsPR2 and OsPR10A protein were increased, implying that these pathogenesis-related proteins were regulated by OsWRKY42 and play roles in Xa21-medicated resistance to Xoo at the downstream of OsWRKY42. In conclusion, the results showed that OsWRKY42 is a novel element in Xa21-mediated resistance pathway against Xoo and enhanced our understanding for the mechanism of rice Xa21-mediated resistance.

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Effects of Overexpression of MtVP1 on Potato Phenotypes and Sugar Metabolism
Jianwu Wang, Wenjuan Wang, Weiwei Xiang, Huiping Dai, Haiqing Wang, Xiangxiang Qu, Furen Kang
Chinese Bulletin of Botany    2022, 57 (2): 197-208.   DOI: 10.11983/CBB21074
Accepted: 13 October 2021

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Type I H+-PPase is involved in gluconeogenesis and sucrose catabolism. Different types of mutants of Arabidopsis thaliana type I H+-PPase gene fed with different sugars (sucrose, glucose and fructose) showed various phenotypes. Therefore, it is speculated that type I H+-PPase may contain other mechanisms that affect sugar metabolism. In order to further clarify the effect of the enzyme on sugar metabolism, phenotypes of the MtVP1 overexpressed potato Weishu 4 were observed under different culture conditions. The fluctuation of sugar content was monitored, and the transcription profile was analyzed. The results showed that the MtVP1 overexpressed potato grew red stem, purple flower, and more developed trichomes. Furthermore, tubers per plant decreased in number, but became larger in size, and shriveled up faster. However, the transgenic plant had significantly decreased starch, glucose, and fructose contents in tubers, and glucose and fructose contents in buds. At the same time, feeding fructose significantly reduced the anthocyanin, and up-regulated the gene expression of fructose-1,6-diphosphate and fructose-2,6-diphosphate by 3-7 fold in the transgenic potato. The results could shed light on further studies of the physiological function of type I H+-PPase from the perspective of sugar metabolism.

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Establishment of Callus Induction and Regeneration System of Anemone silvestris
Jinchun Lu, Lina Cao, Guanjie Tong, Xinying Wang, Liying Zhang, Xin Yu, Huifang Li, Yanhui Li
Chinese Bulletin of Botany    2022, 57 (2): 217-226.   DOI: 10.11983/CBB21200
Accepted: 04 January 2022

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In order to establish the tissue culture and regeneration system of wild Anemone silvestris, we investigated effects of different concentrations of plant growth regulators on callus induction, adventitious bud differentiation, proliferation and rooting of different explants including epicotyl, hypocotyl, leaf and petiole. The results showed that adventitious buds could be induced in all four explants, among them the epicotyl was the best. The best medium for hypocotyl induction was 1/2MS+2.0 mg∙L-1 6-BA+0.1 mg∙L-1 NAA with the highest induction rate of 86.67%; The optimal medium for proliferation was as follow 1/2MS+1.0 mg∙L-1 6-BA+0.05 mg∙L-1 NAA, and the proliferation coefficient was 3.67; The optimal rooting medium was 1/2MS+0.3 mg∙L-1 IBA with the rooting rate of 100%; In the medium of peat:vermiculite = 2:1 (v/v), the survival rate of tissue culture seedling was at 98.33%. This study effectively solved the shortage of germplasm resources of A. silvestris in garden and medicinal production, and provided technical support for the realization of factory seedling cultivation.

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Advances in the Regulation of Rice (Oryza sativa) Grain Shape by Auxin Metabolism, Transport and Signal Transduction
Lixia Jia, Yanhua Qi
Chinese Bulletin of Botany    2022, 57 (3): 263-275.   DOI: 10.11983/CBB21227
Accepted: 18 March 2022

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Rice (Oryza sativa) is a major food crop in the world. The optimization and utilization of the major yield-determining factors are important for increasing yield potential. Among these factors, seed weight is one of the most important factors determining rice production. The heritability of rice grain weight is stable, which is largely unaffected by environmental factors. Grain weight depends on grain shape, which is determined by grain length, grain width, and grain thickness, and the degree of grain filling. The growth of rice glumes and seed endosperm determines the grain shape and weight. The proliferation and expansion of glume cells affect grain development, and endosperm occupies most of the volume of mature seeds. Auxin is an important plant hormone that affects rice yield, which regulates the development of glume and endosperm after fertilization. The spatial-temporal distribution of active auxin is dynamically modulated by auxin metabolism, auxin transport and signal transduction, all of which maintain auxin at the optimal level for seed development. Here we reviewed the research progress of auxin pathways regulating rice grain shape from three aspects, auxin metabolism, auxin transport and auxin signal transduction, to provide clues for exploring the auxin regulation mechanism of grain shape and improve yield in rice.

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The Measurement Principles, Methods and Applications of P515
Chunyan Zhang, Xiaojie Pang
Chinese Bulletin of Botany    2021, 56 (5): 594-604.   DOI: 10.11983/CBB21052
Accepted: 18 June 2021

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The spectral techniques have been widely used in the field of photosynthesis research, such as the light absorption signals P515 and P700 redox kinetics, and chlorophyll fluorescence, which can detect the photosynthetic activities of plants quickly and accurately. P515 signal is widely present in higher plants and algae, which is caused by the shift of absorption spectrum of pigments on thylakoid membrane. We can detect the ratio of PSI to PSII reaction center, the proton conductivity of chloroplast ATP synthase, the cyclic electron flow rate around PSI, the proton motive force (pmf) and its components by the P515 fast and slow kinetics, and study the photoprotective mechanism by simultaneous detection of P515 signal and chlorophyll fluorescence. In this paper, we summarize the main measurement methods of P515, expound its principles, and the applications. The aim is to provide technical supports for further study on the mechanism of photosynthesis.

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Green Extraction Method and Optimization of Steviosides from Stevia rebaudiana by Natural Deep Eutectic Solvent
Qing Miao, Pa-he-er-ding Sabiha, Siyu Zeng, Qifang Pan
Chinese Bulletin of Botany    2021, 56 (6): 722-731.   DOI: 10.11983/CBB21064
Accepted: 17 September 2021

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Natural deep eutectic solvent (NADES) was used to extract steviosides from Stevia rebaudiana as an efficient, green and environmentally friendly new method. Compared with traditional solvent water, the optimal NADES formula was selected to extract S. rebaudiana dry leaves by measuring the concentration of stevioside and rebaudioside A, two main components in steviosides. The extraction conditions of NADES extracting steviosides were optimized by Box-Behnken design from response surface methodology. The best NADES formula for steviosides extraction was selected: 1,2-propanediol:glycerol:water=8:1:1 (v/v/v), the extraction concentration of stevioside was 2.59 mg∙mL-1 (16.40% higher than water), and the extraction concentration of rebaudioside A was 1.06 mg∙mL-1 (12.62% higher than water). The NADES extraction conditions were optimized by response surface methodology: extraction time was 90 min, extraction temperature was 60°C, ultrasonic power was 80 J∙s-1, the predicted extraction concentration of stevioside was 3.49 mg∙mL-1 and the concentration of rebaudioside A was 1.43 mg∙mL-1, which was close to the experimental verification value (the concentration of stevioside is 3.48 mg∙mL-1, the concentration of rebaudioside A is 1.42 mg∙mL-1). Under the best condition, the extraction of stevioside was 34.36% higher than the initial condition, and the extraction of rebaudioside A was 33.96% higher than the initial condition. NADES is environmentally friendly and has higher extraction efficiency than traditional solvents. It can be used for the green extraction of steviosides from S. rebaudiana. At the same time, it provides a promising prospect for the green extraction of natural products from other bulk economic plants.

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Seed Development and Embryo Culture of Endangered Abies beshanzuensis
Ke Liu, Bin Liu, Lu Yuan, Hui Shuai, Yang Yang, Tingjin Wang, Deliang Chen, Xiaorong Chen, Kaibin Yang, Xiaofeng Jin, Liping Chen
Chinese Bulletin of Botany    2021, 56 (5): 573-583.   DOI: 10.11983/CBB21096
Accepted: 12 October 2021

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The seed germination rate of Abies beshanzuensis is low, and the characteristics of seed development are elusive, which severely limit the natural regeneration of the population in situ. To illustrate the developmental characteris-tics of seeds and identify the main factors affecting seed development, this article aimed to study the number, weight, and microstructural characteristics of the embryo and endosperm of the seed in the female cones at different development stages, as well as the primary metabolites of the endosperm in the critical period of seed development. The results showed that July was a critical period for seed development of A. beshanzuensis. During this period, the endosperm weight increased rapidly and the carbohydrate metabolism in the endosperm was active. Anatomical and morphological analysis of the female cones showed that a large number of abnormally developed seeds appeared in cones in late July. Analysis of the primary metabolites of endosperm during the critical period showed that the content of trehalose-6-phosp-hate in the endosperm of abnormally developed seeds was significantly decreased. It was speculated that during the critical period of seed development, the decrease of trehalose-6-phosphate in the endosperm may be an important cause of abnormal seed development. Based on the characteristics of A. beshanzuensis seed development, we established the embryo culture technology for A. beshanzuensis, and generated test-tube plantlet.

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Effects of Exogenous Melatonin on Physiological Response and DNA Damage of Ardisia mamillata and A. crenata Under Lead Stress
Jinxiang Ai, Jiayi Song, Zhenan Yan, Zhichao Wang, Wenqian Chen, Yuhuan Wu, Yanyan Wang, Leilei Pan, Yutao Xu, Peng Liu
Chinese Bulletin of Botany    2022, 57 (2): 171-181.   DOI: 10.11983/CBB21191
Accepted: 07 February 2022

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Lead (Pb) is a major abiotic constraint affecting plant growth. To compare the anti-lead ability of two Ardisiaspecies and investigate the physiological responses and DNA damage to the treatments of exogenous melatonin (MT) under lead stress, hydroponic experiment was carried out with 2-year-old seedlings of A. mamillata and A. crenata. The results showed that with the increasing of the lead stress concentration in the same period of time, the three antioxidant enzyme activities, proline (Pro) and soluble protein (SP) contents of the A. mamillata and A. crenata were all increased first and then decreased, while malondialdehyde (MDA) content decreased at first and then increased, the root tip callosin content continues to rise, DNA damage in roots was increased. After the application of appropriate concentrations of exogenous MT, the activities of antioxidant enzymes in both plants were effectively enhanced and reached a maximum at 100 µmol∙L-1 MT treatment under different concentrations of Pb, and the contents of Pro and SP were significantly increased. However, the MDA content decreased significantly at first and then increased slowly. The root tip callosin content continues to increase and the root DNA damage was improved. With the increasing of MT concentration, the alleviating effect of melatonin on A. mamillata and A. crenata gradually weakened. A. crenata is more resistant to lead stress and its physiological response is more stable than A. mamillata. Exogenous application of MT can effectively alleviate the toxic effect of lead stress on A. mamillata and A. crenata (relief effect of A. crenata > A. mamillata), and enhance its tolerance to lead toxicity, among which 100 µmol∙L-1 MT treatment has the best mitigation effect. We revealed the superiority of lead resistance of A. mamillata and A. crenata, and the alleviating effect of exogenous MT on lead poisoning, thus providing a theoretical reference for the study of lead resistance of Ardisia.

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