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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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Current Issue
Editor-in-Chief:Kang Zhong
ISSN 1674-3466 CN 11-5705/Q
Post Code:2-967
Volume 58 Issue 6
01 November 2023
  
COMMENTARIES
New Insights Into the Origin of Modern Maize-hybridization of Two Teosintes
Xiting Yu, Xuehui Huang
Chinese Bulletin of Botany. 2023, 58(6):  857-860.  doi:10.11983/CBB23138
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The domestication of crops was a significant event in human history, which led to the emergence and prosperity of agricultural civilization. Maize is an important global food crop, and its domestication origin has long attracted the attention of both the biological and historical communities. The mainstream view in the past was that modern maize originated from the parviglumis type of teosinte. Recently, Yan Jianbing and his collaborators systematically collected and sorted various types of wild and cultivated maize resources, and comprehensively applied genomics, population genetics, and quantitative genetics methods, along with the use of archaeological findings. They found that modern maize also has the gene introgression of the mexicana type of teosinte, which has influenced many agronomic traits. A new model for the origin of modern maize has been proposed based on these findings.

There Is a Way Out-new Breakthroughs in Extracellular Auxin Sensing
Xiangpei Kong, Mengyue Zhang, Zhaojun Ding
Chinese Bulletin of Botany. 2023, 58(6):  861-865.  doi:10.11983/CBB23149
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Auxin plays an important role in plant growth and development and its signal transduction has always been the focus of attention in the field of plant biology. AUXIN BINDING PROTEIN 1 (ABP1)-TRANSMEMBRANE KINASE (TMK) molecular module is involved in the extracellular auxin perception. In recent years, ABP1 has been controversial as an auxin receptor. Recently, Tongda Xu’s team and Zhenbiao Yang’s team from Fujian Agriculture and Forestry University identified ABP1-LIKE PROTEIN (ABL) as the auxin binding proteins involved in the extracellular auxin perception. Different from traditional functional redundancy, ABL and ABP1 achieve functional compensation effect through protein structure similarity, and then form complex with TMK at the plasma membrane, acting as co-receptors of apoplastic auxin to mediate auxin driven rapid response. This study deeply dissects the mechanism of extracellular auxin sensing, which is a breakthrough in the field of auxin signaling transduction.

EXPERIMENTAL COMMUNICATIONS
ZmICE2 Regulates Stomatal Development in Maize
Wenqi Zhou, Yuqian Zhou, Yongsheng Li, Haijun He, Yanzhong Yang, Xiaojuan Wang, Xiaorong Lian, Zhongxiang Liu, Zhubing Hu
Chinese Bulletin of Botany. 2023, 58(6):  866-881.  doi:10.11983/CBB22261
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Plant epidermis is crucial in regulating photosynthesis, respiration, heat dissipation, and water utilization. Significant progress has been made in the study of stomatal development in dicotyledonous plants, such as Arabidopsis thaliana. Three important bHLH transcription factors (SPCH, MUTE, and FAMA) have been reported to be specifically expressed at different stages of cell division and differentiation in the stomatal lineage. They form heterodimers with another transcription factors SCRM/ICE1 and SCRM2/ICE2 to regulate the morphological transformation and changes of stomatal lineage cells across three stages of division, finally forming the stomatal complex. However, in monocots, especially in Poaceae plants such as maize (Zea mays), studies on genes regulating epidermal morphogenesis are less reported. In this study, two single-gene recessive mutants, Zmice1-1 (inducer of cbf expression1-1) and Zmice2-1, were isolated using reverse genetics approaches. Compared to the control B73, Zmice2-1 exhibited dwarfism, leaf chlorosis, reduced fertility, significantly lower stomatal density and index, disrupted arrangement of epidermal long cells, and absence of spacing between stomata. Zmice1-1 leaves gradually turned yellow from the five-leaf stage and displayed complete chlorosis at later stages. The homozygous Zmice1-1 plants are growth-arrested and sterile, but the stomatal density showed no significant difference compared to the control. Different allels of Zmice2 were obtained using CRISPR-Cas9 genome editing technology. Phenotypic identification showed that Zmice2-2 had an abnormal stomatal phenotype similar to Zmice2-1, indicating that ZmICE2 is involved in the regulation of stomatal development. Transcriptome analysis of B73 and Zmice2-1 revealed that ZmICE2 primarily regulated stomatal development by affecting cell division and differentiation, participating in the formation of maize epidermal morphology. These results contribute to a better understanding of the mechanisms of epidermal morphogenesis in maize and provide valuable genetic resources for improving crop resilience and yield traits.

Mapping of QTL for Cell Wall Related Components in Rice Stem and Analysis of Candidate Genes
Qiwei Jia, Qianqian Zhong, Yujia Gu, Tianqi Lu, Wei Li, Shuai Yang, Chaoyu Zhu, Chengxiang Hu, Sanfeng Li, Yuexing Wang, Yuchun Rao
Chinese Bulletin of Botany. 2023, 58(6):  882-892.  doi:10.11983/CBB23100
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Lodging of rice is one of the main factors reducing rice yield. The mechanical strength of stem affects lodging resistance of rice and is closely related to the content of stem cell wall related components. Improving lodging resistance of rice by regulating the related components in the cell wall of stem is an effective way to improve the yield and quality of rice. In this study, indica rice variety Huazhan (Oryza sativa subsp. indica cv. ‘HZ’) and japonica rice variety Nekken2 (O. sativa subsp. japonica cv. ‘Nekken2’) were crossed to obtain F1 generation. A total of 120 RILs (recombinant inbred lines) population were obtained by successive multigeneration self-crossing and the genetic linkage map was constructed. Based on the constructed high-density genetic map, the QTLs related to the content of cellulose, hemicellulose, and lignin in the cell wall of rice stem were located and analyzed. The results showed that 4 QTLs related to cellulose, 12 QTLs related to hemicellulose, and 8 QTLs related to lignin. At the same time, candidate genes analyses were conducted on the detected QTLs’ intervals, and a total of 13 candidate genes were screened. The expression levels of candidate genes were detected by qRT-PCR. Except for LOC_Os02g58590 and LOC_Os12g41720, the other candidate genes showed significant differences between parents. The results laid an important foundation for exploring genes that regulate the mechanical strength of rice stems, further screening and breeding rice varieties with strong lodging resistance.

Morphological Identification and Development of Linkage Markers for Lobed Ray Floret Mutants in Marigold (Tagetes erecta)
Wenjing Wang, Yu Zhu, Hongming Zhang, Ludan Wei, Qingping Yi, Xiaomin Yu, Yuhan Liu, Lixue Zhang, Wenhan Cheng, Yanhong He
Chinese Bulletin of Botany. 2023, 58(6):  893-904.  doi:10.11983/CBB22231
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Marigold (Tagetes erecta) is an important ornamental plant of the genus Tagetes in the Asteraceae. The morphology of its ray floret corolla varies from smooth, undulate to different degree of incision. An F2 population was constructed from a cross between the mutant marigold JH with five-lobed corollas and fertile stamens in the first round florets and the inbred line S5 with smooth corolla of ray florets and aborting stamens. Inheritance analysis showed that the mutation trait of JH was controlled by a single-dominant gene, named Tagetes erecta lobed ray florets (Telf). Histological and cytological observations showed that the first round ray florets of JH were disk-like, with five lobes formed at the top of the corolla tube, the middle and lower parts were tubular, and the development of anthers and pollen grains are to normal. Using BSR-seq and comparative genomics methods, the gene Telf controlling lobed ray florets was mapped in the interval of 37003-SCAR and 34032-CAPS. 37003-SCAR marker was 3.684 cM from the Telf gene. On the other side of the gene, 34032-CAPS marker was placed about 3.517 cM from Telf. This study laid a foundation for the subsequent fine mapping of the target gene Telf, and also provided a new method for the selection of molecular markers in marigold.

TECHNIQUES AND METHODS
Establishment of CRISPR/Cas9 Gene Editing System in Rehmannia henryi
Chunyan Miao, Mingming Li, Xin Zuo, Ning Ding, Jiafang Du, Juan Li, Zhongyi Zhang, Fengqing Wang
Chinese Bulletin of Botany. 2023, 58(6):  905-916.  doi:10.11983/CBB22250
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Rehmannia henryi is an important plant with great medicinal value, but no research of CRISPR/Cas9 has been done on this species. To establish the gene editing system of R. henryi, the gene encoding phytoene desaturase (PDS) in carotenoid biosynthesis was screened, and the CRISPR/Cas9 vector of RhPDS1 was constructed and transformed into R. henryi genome by Agrobacterium-mediated transformation method. The transcript of RhPDS1 with a 1 764 bp open reading frame (ORF) of RhPDS1 was obtained, the deduced amino acid sequence of RhPDS1 has the typical structural domains of phytoene desaturase. RhPDS1 showed higher expression levels in bud, flower and new leaf. Using CRISPR/Cas9 method, three regenerated shoots with albino phenotype were finally obtained, the differentiation rate of albino shoot was 3.7%. Sequencing analysis revealed that the three albino shoots belong to 2 editing events, in which deletion of 1 bp or (and) 5 bp occurred, respectively, which caused frame shift mutations. The contents of chlorophylls and carotenoids were significantly decreased in the albino mutants as compared to wild type, and the expression levels of RhPDS1 were also decreased in the albino mutants. Taken together, the RhPDS1 gene was cloned and knocked out by using CRISPR/Cas9 method, which laid down the foundations for functional genomics studies and de novo domestication of R. henryi.

In vitro Culture of Taxus Rich in Taxanes
Xiaohui Feng, Xuetong Yan, Keyuan Zheng, Qiang Zhou, Weizhong Zhang, Quanyong Wang, Mulan Zhu
Chinese Bulletin of Botany. 2023, 58(6):  917-925.  doi:10.11983/CBB22228
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Taxus contains the anti-cancer ingredient paclitaxel, is the top-protected endangered plant in China, and has great medicinal value but scarce resources. To solve the problem of paclitaxel source shortage, in this study, an in vitro culture of a taxane-rich Taxus system was established. The results showed that MS+2 mg·L-1 6-BA+0.4 mg·L-1 NAA+0.7 g·L-1 Pro was the optimal treatment for absorption surface expansion, and the expansion rate reached 90%. The adventitious bud high-frequency synchronous growth induction was in the DCR+1 mg·L-1 6-BA+0.1 mg·L-1 NAA+5 mg·L-1 Glu+1 g·L-1 AC. The best biomass amplification phase I was in MS+0.7 mg·L-1 6-BA+0.07 mg·L-1 NAA+0.1 mg·L-1 Phe+50 mg·L-1 PG, in which the biomass reached 542 mg. The optimized biomass amplification phase II was obtained in MS+0.5 mg·L-1 6-BA+0.05 mg·L-1 NAA+0.1 mg·L-1 Phe+2 g·L-1 AC, in which the biomass reached 1 612 mg. The content of taxanes in tissue culture materials is much higher than that of natural materials. The content of paclitaxel in tissue culture materials was 6.1 times that of natural materials. The contoent of baccatin III in tissue culture materials was 8.2 times that of natural materials. The content of 10-DAB in tissue culture materials was 68.1 times that of natural materials. This study established an in vitro culture system of taxane-rich Taxus, and solved the problem of paclitaxel drug shortage.

Establishment of In Vitro Regeneration System of Citrus australasica
Chungang Xie, Zhe Liu, Shusheng Zhang, Haitao Hu
Chinese Bulletin of Botany. 2023, 58(6):  926-934.  doi:10.11983/CBB23060
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In order to establish the regeneration system of Citrus australasica, the effects of different plant growth regulators combinations, medium types and dark culture time on callus induction and plant regeneration of C. australasica were studied using stem segments as explants. The results showed that the best medium for adventitious bud induction was 1/2MS+4.0 mg∙L-1 ZT+30.0 g∙L-1 sucrose. Dark culture for 14 days and then light culture had the best promoting effect. The induction rate of callus and adventitious bud was 100%, and the average number of adventitious bud regeneration per explants was 4.83. The optimal rooting medium was 1/2MS+0.5 mg∙L-1 NAA, and the regenerated plants with the 94.43% rooting rate were obtained, and the average number of roots was 3.9. In the mixture of grass carbon:perlite: vermiculite=2:1:1 (v/v/v), tissue culture seedlings had the best growth, and the survival rate was more than 90%. This study established in vitro regeneration system of C. australasica, which laid the foundation for the genetic improvement and rapid propagation of C. australasica fine varieties.

SPECIAL TOPICS
Aptamers and Their Applications in Plant Science Researches
Yi Li, Xi Zhang, Yanhui Yuan, Pichang Gong, Jinxing Lin
Chinese Bulletin of Botany. 2023, 58(6):  935-945.  doi:10.11983/CBB23050
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Aptamers are single-stranded oligonucleotides or short peptides that are screened and selected from artificially synthesized libraries and can specifically bind to the target molecules. Aptamers can bind and regulate the activities of its target molecules based on their own structure and sequence, and are applied to in vivo molecular function researches and new drug preparation explorations. In recent years, peptide aptamers have been widely used in various fields, such as medicine, genetics and molecular biology, which is becoming a highly efficient, specific and powerful new tool. In plants, with the establishment, application and promotion of the corresponding system, aptamers have gradually been become an effective tool for studying plant molecular functions. In this review, we summarized the classification, screening principles and basic applications of aptamers, with special emphasis on their applications in plant breeding. It can be expected that with the development of molecular design bio-breeding technology, aptamers can be cooperated with transgenic techniques and become a valuable application tool in the plant science fields.

Advances in the Regulation of Protein Liquid-liquid Phase Separation on Development and Stress Responses in Plants
Xinhua Huang, Wei Liu, Shiping Tian, Tong Chen
Chinese Bulletin of Botany. 2023, 58(6):  946-955.  doi:10.11983/CBB23070
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Cells are basic units of structure and function for organisms, in which membranous and membraneless organelles exert their respective functions in a given space. However, the mechanism underlying the formation and function of membraneless organelles still requires further investigation. It has been shown that liquid-liquid phase separation (LLPS) of proteins may provide a specific way for biomacromolecules within cells to aggregate and form agglomerates, then proceed to play important roles. With increasing reports on LLPS in plants, great interest has been in exploring the roles of phase separation-forming agglutinates in plant growth and development and immune processes. In this review, we mainly focus on the progress of current studies on the role of protein liquid-liquid phase separation in regulating plant growth and development as well as stress responses, which may provide references for further studies on the mechanism of LLPS in plant cells.

The Application of Click Chemistry Reactions in Plant Cell Labeling
Yuge Zhang, Xiaoyan Yuan, Guifang Zhang, Yujian Li, Jinhuan Yin, Jinxing Lin, Xiaojuan Li
Chinese Bulletin of Botany. 2023, 58(6):  956-965.  doi:10.11983/CBB22252
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Click chemistry, also known as "link chemistry" or "speed-matching combinatorial chemistry", is the development of a powerful, highly reliable, and highly specific set of reactions for the rapid synthesis of new compounds through the connection of carbon-heteroatomic bonds (C-X-C). Click chemistry has been progressing greatly in drug development, new material synthesis, material surface functionalization modification and biological macromolecular labeling. Pioneers of click chemistry was awarded the Nobel Prize in Chemistry in 2022. This review briefly introduces the principles, reaction types and applications of click chemistry, summarizes the research progress of click chemistry in labeling biological macromolecules, especially in its application to plant cell wall polysaccharide labeling, and provides new ideas for the study of plant cell wall synthesis, structure and dynamic transport.

Research Progress in Plant Antifreeze Protein
Xiaoyun Dong, Jiaping Wei, Junmei Cui, Zefeng Wu, Guoqiang Zheng, Hui Li, Ying Wang, Haiyan Tian, Zigang Liu
Chinese Bulletin of Botany. 2023, 58(6):  966-981.  doi:10.11983/CBB22248
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Temperature is one of the major environmental determinants of plant geographical distribution. Plants distri-

buted at high latitudes or altitudes usually suffer a period of sub-zero temperature during their life cycle. When the ambient temperature drops to the freezing point, the water molecules in plants form ice crystals, causing fatal damage to plant tissue structure. In order to adapt to the freezing environment, the pathogenesis-related protein PR (PR) and its related WRKY transcription factors in cold climate plants have evolved into antifreeze proteins (AFPs) that can bind specifically to the ice surface and effectively inhibit the formation and growth of ice crystals. Currently, AFPs have been identified in nearly 100 plant species, such as winter rye (Secale cereale). Compared with insect AFPs, plant AFPs have extremely high recrystallization inhibition activity, which can effectively prevent the formation of large ice crystals in vivo. Both low temperature and pathogens can induce AFP synthesis in cold climate plants. Interestingly, only the cold-induced AFPs have dual molecular functions as hydrolase/antifreeze activity. The PR-AFPs, however, possess only one of hydrolase/antifreeze activites, whose conversion is controlled/regulated by post-translational peptide differential folding, as suggested with growing evidence. AFP has gradually become one of the hot targets in botanical research due to its unique molecular function and its promising applications. This paper will provide a systematic review of recent progress in this area.

Research Advances in Biological Functions of Plant SPL Transcription Factors
Xinhai Zeng, Rui Chen, Yu Shi, Chaoyue Gai, Kai Fan, Zhaowei Li
Chinese Bulletin of Botany. 2023, 58(6):  982-997.  doi:10.11983/CBB22216
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Squamosa promoter binding protein-like (SPL) family is a class of plant-specific transcription factors, which contain a highly conserved SBP domain consisting of two zinc finger structures and a short nuclear localization sequence. The expression of most SPL genes is regulated by microRNAs at transcription level. Based on the current research progress of SPL transcription factors, this paper summarizes the biological functions of SPLs in plant growth, development, and environmental adaptation, and discusses the future research directions of SPLs.

Research Progress in Sirtuin Protein Family in Plants
Lufang Su, Ping Wang, Shun Li, Yan Cai, Dandan Guo, Qin Liu, Xiaoyun Liu
Chinese Bulletin of Botany. 2023, 58(6):  998-1007.  doi:10.11983/CBB22255
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The sirtuin protein family of plants is a class of enzymes that depend on β-nicotinamide adenine dinucleotide (β-NAD+) to catalyze the removal of Nε-acetyl and Nε-acyl (acetyl, crotonyl, butylyl and 2-hydroxy-isobutylyl) from non-histone and histone lysine, respectively, and has the function of regulating the activity of non-histone protein and gene expression. Members of the sirtuin protein family have been shown to play important epigentic regulatory roles in the growth and development of rice (Oryza sativa), soybean (Glycine max) and tomato (Solanum lycopersicum), and in response to salt, cold, heat and pathogen stresses. Here we summarized the recent research advances in the enzyme activity, catalytic substrates, cell localization and function of plant sirtuins, which will serve as a reference for understanding the epigenetic regulation mechanism and studying the new function of sirtuin.

Research Progress on Physiological Functions of Suberin lamellae in Water and Solutes Transport
Biao Zhang, Jian Wu, Yang Zhang, Xiaowei Dong, Shuo Han, Xin Gao, Congwu Du, Huiying Li, Xuefa Chong, Yingying Zhu, Haiwei Liu
Chinese Bulletin of Botany. 2023, 58(6):  1008-1018.  doi:10.11983/CBB22208
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Plant roots can acquire water and nutrients selectively from soil and transport them upwards to the aerial parts for plant growth and development. These functions are closely related to their anatomical structures. The radial transport of water and solutes absorbed by roots mainly includes three different pathways, namely, the apoplastic pathway, the symplastic pathway and the transcellular pathway. The endodermis is the innermost cell layer that surrounds the central vasculature. For a long time, endodermal differentiation formed the Casparian strips has been considered to play a decisive role in blocking water and solutes transport through the apoplastic pathway. However, in recent years, it has been found that suberin lamellae formed by endodermal differentiation plays no less important role in the radial transport of water and solutes than Casparian strips, and even that suberization is the second life of an endodermal cell. In this paper, we reviewed the latest research progress on the physiological function of suberin lamellae in water and solutes transport in recent years, and discussed the relationship between suberin lamellae and drought, salt, nutrient and heavy metal stress of crops, in order to provide a reference for the theory and practice of endodermal plasticity in the regulation of plant physiological function.

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