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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 55 Issue 1
01 January 2020
  
COMMENTARIES
The Development of Genomics Technologies Drives New Progress in Horticultural Plant Research
Tang Jiali,Qiu Jie,Huang Xuehui
Chin Bull Bot. 2020, 55(1):  1-4.  doi:10.11983/CBB19240
Abstract ( 253 )   HTML ( 5 )   PDF (802KB) ( 346 )   Save
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Horticultural plants include flowers, vegetables, fruit trees, some melon (such as watermelon (Citrullus lanatus), muskmelon (Cucumis melo)) and tea trees (Camellia sinensis), with a large number of species based on plant classification. Genomics and genetics researches for horticultural plants are of important theoretical value and economic significance. The development of genome sequencing technology and related bioinformatics tools greatly facilitate molecular biology researches of horticultural plants. In addition to its important ornamental value, the plant species ‘water lily’ has a very special position in evolutionary, belonging to an early angiosperm group. Recently, a high-quality genome map of an important flower plant, water lily, was generated. Through systematic analysis and genomic comparison of the water lily genome and other angiosperm genomes, the researchers thoroughly elucidated the evolutionary position and related evolutionary events of water lily. Based on the high-quality genomic sequences of these horticultural plants, researchers in horticultural plant science are expected to carry out in-depth molecular genetics research and identify functional genes underlying many traits such as flower organs, flower color, fragrance, and quality, which is expected to promote basic research and accelerate the creation of new varieties.

A New Progress of Green Revolution: Epigenetic Modification Dual-regulated by Gibberellin and Nitrogen Supply Contributes to Breeding of High Yield and Nitrogen Use Efficiency Rice
Han Mei-ling,Tan Ru-jiao,Chao Dai-yin
Chin Bull Bot. 2020, 55(1):  5-8.  doi:10.11983/CBB20002
Abstract ( 412 )   HTML ( 2 )   PDF (1225KB) ( 324 )   Save
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The Green Revolution represented by the breeding of semi-dwarf crops greatly promoted agriculture yield, but it also unfortunately led to the problem of low nitrogen use efficiency (NUE). The achievement of Green Revolution was mainly based on modification of gibberellin (GA) metabolic or signaling pathways in crops. A previous study has found that the central regulator of GA signaling pathway DELLA protein negatively regulates NUE through suppressing GRF4, an essential NUE regulator, which provided a resolution for improving NUE of semi-dwarf rice. A recent study further revealed a novel mechanism underlying the crosstalk between GA signaling and nitrogen response. The study revealed that NGR5 is a key gene controlling tiller number changes under different nitrogen conditions, which is inducible by nitrogen. Further investigation established that the NGR5 suppresses branching inhibitory genes, such as D14 and OsSPL14, through nitrogen-dependent recruitment of polycomb repressive complex 2 that promotes histone H3 lysine 27 tri-methylation in the regions habouring the branching suppressors. In addition to be responsive to nitrogen, NGR5 is also negatively regulated by GA and its receptor GID, and overexpression of NGR5 in the semi-dwarf background is thus able to significantly improve rice yields under low nitrogen conditions. This study not only uncovered a new mechanism of GA signaling, but also enlightens the new generation of Green Revolution by breeding high yield crops with enhanced NUE.

EXPERIMENTAL COMMUNICATIONS
Transformation of Insect Derived Antifreeze Gene into Sweet Potato (Ipomoea batatas) and Enhanced Its Freeze-tolerance
Lai Xianjun,Zhang Yizheng,Gu Yinghong,Yan Lang
Chin Bull Bot. 2020, 55(1):  9-20.  doi:10.11983/CBB19133
Abstract ( 160 )   HTML ( 2 )   PDF (14013KB) ( 114 )   Save
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To explore whether the gene encoding antifreeze protein from insect can enhance the freezing tolerance of sweet potato through gene transformation, and to prepare freeze-tolerance materials for breeding purposes, we constructed a plant gene expression vector harboring an antifreeze gene TmAFP from yellow mealworms (Tenebrio molitor) and obtained transgenic freeze-tolerance sweet potato lines using Agrobacterium-mediated transformation method. A high-frequency regeneration system of sweet potato was established using the variety Huachano as the recipient material, and the embryogenic suspension cells were cultured in the somatic embryo maturation medium. The sensitivity test of embryogenic cells to herbicides indicated that the combination of MS+0.2 mg·L -12,4-D+0.8 mg·L -1GAP+100 mg·L -1Carb is the most effective medium for screening the transgenic positive plants. Seven embryogenic calli were obtained and 42 resistant seedlings were regenerated, among which 23 harbored pSUIBEV3-AFP and 19 had pCAMBIA-AFP. All resistant seedlings were examined by PCR, Southern hybridization and RT-PCR, and the results showed that the TmAFP gene was integrated into the plant genome and expressed. The transgenic and non-transgenic plants were treated at -1°C for 15 hours, and then transferred to room temperature. The results demonstrated that the freeze-tolerance of the transgenic plants was greatly improved.

Effect of Exogenous Substances on Cold Tolerance and Key Sucrose Metabolic Gene Expression in Camellia sinensis
Yang Xiaoqing,Huang Xiaoqin,Han Xiaoyang,Liu Tengfei,Yue Xiaowei,Yi Ran
Chin Bull Bot. 2020, 55(1):  21-30.  doi:10.11983/CBB19024
Abstract ( 362 )   HTML ( 2 )   PDF (1346KB) ( 132 )   Save
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In recent years, low temperature damage in winter and spring has become an important factor affecting tea production. We used tea tree (Camellia sinensis) potted seedlings to test the effect of GABA, chlorella powder and bamboo vinegar on sucrose-related gene expression (SPS, SUS4, INV4, INV5) and cold-tolerant related physiological indexes. Different concentrations of the three kinds of exogenous substances were sprayed on seedlings under cold stress. We analyzed the expression of SPS, SUS4, INV4 and INV5 of the key sucrose-coding genes as well as the physiological indexes related to cold tolerance to determine the possible physiological and molecular mechanisms of the effect of exogenous substance spraying on cold tolerance of tea trees. The frost index and relative conductivity were significantly lower after application of GABA, chlorella powder or bamboo vinegar than control values and the soluble sugar content was increased. The optimal application concentration for GABA, chlorella powder and bamboo vinegar was 10 mmol·L -1, 0.22 mg·mL -1 and 2.5 mg·mL -1, respectively. Malondialdehyde content was significantly decreased and antioxidant enzyme activity was increased; chlorophyll, soluble sugar and proline content was also increased. Sucrose content was increased by 15.24%, 11.39% and 5.97%, respectively, after 72 h treatment. The expression of SPS, SUS4, INV4 and INV5 in leaves was significantly increased with 10 mmol·L -1 GABA, 0.22 mg·mL -1 chlorella powder and 2.5 mg·mL -1 bamboo vinegar. Thus, GABA, chlorophyll powder and bamboo vinegar can significantly improve the cold tolerance of tea plants. We provide a theoretical basis for the cryoprotectant selection for tea plants.

Effects of the Respiratory Electron Transport Pathways in Relieving Photoinhibition of Chloroplast PSII in Tobacco Leaves
Luo Jiao,Li Yuting,Zhang Zishan,Che Xingkai,Liang Ying,Li Yuenan,Li Ying,Zhao Shijie,Gao Huiyuan
Chin Bull Bot. 2020, 55(1):  31-37.  doi:10.11983/CBB19117
Abstract ( 108 )   HTML ( 3 )   PDF (1401KB) ( 80 )   Save
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Previous studies have shown that the mitochondrial alternative oxidase (AOX) respiratory pathway contributes to the photoprotection of photosystem II (PSII) under high light. However, the role of another respiratory pathway in mitochondria, the cytochrome-respiratory (COX) pathway, to photoprotection under high light remains unknown. Here, we studied the contributions of COX and AOX pathways to PSII photoprotection, as well as the relationship between COX and AOX pathways in PSII photoprotection in tobacco leaves under high light using chlorophyll fluorescence transients and chlorophyll fluorescence quenching approaches. After high light treatment, the PSII activity decreased in all leaves. The inhibition of AOX pathway significantly accelerated the decrease of PSII activity in leaves. However, there was no significant difference between the PSII activity in the leaves pretreated with COX pathway inhibitor and water-pretreated control leaves. When both AOX and COX pathways are inhibited in leaves, the decrease in PSII activity was severe, than that of leaves pretreated by only AOX pathway inhibitor. In addition, inhibition of respiratory electron transport resulted in an increase of non-photochemical quenching (NPQ). The up-regulation of NPQ caused by AOX pathway inhibitor was more obviously than that caused by COX pathway inhibitor, and the increase of NPQ was highest when both AOX and COX pathways were inhibited. These results indicate that both COX and AOX pathways contribute to the PSII photoprotection in tobacco leaves under high light. In addition, when COX pathway is inhibited, its photoprotective function can be compensated by AOX pathway and NPQ, however, photoprotection of AOX pathway cannot be completely compensated by COX pathway and NPQ.

Expression Patterns and Bioinformatic Analyses of Salt Stress Responsive Gene IbMYB3 in Ipomoea batatas
Li Ge,Meng Xiaoqing,Li Zongyun,Zhu Mingku
Chin Bull Bot. 2020, 55(1):  38-48.  doi:10.11983/CBB19094
Abstract ( 160 )   HTML ( 2 )   PDF (8533KB) ( 109 )   Save
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MYB transcription factors have multiple biological functions and play important roles in mediating plant responses to biotic and abiotic stresses. In this paper, two MYB genes, named IbMYB3 and IbMYB4, which were significantly induced by salt stress, were screened from RNA-seq data of salt-stressed sweetpotato (Ipomoea batatas) plantlets. Gene expression analysis showed that the expression of IbMYB3 was significantly up-regulated by various abiotic stresses and plant growth substances treatments, suggesting that IbMYB3 might be involved in the abiotic stress responses of sweetpotato. Further bioinformatic analysis showed that the open reading frame of IbMYB3 is 1 059 bp in length, encoding 353 amino acids, with a predicted molecular weight of 39.41 kDa and the theoretical isoelectric point (PI) of 5.26, which is an acidic negatively charged hydrophilic protein. Subcellular localization showed that the IbMYB3 protein localizes to the nucleus, and has strong transcriptional activation activity. Taken together, these results demonstrated that the IbMYB3 transcription factor might play an important role in regulating the abiotic stress responses of sweetpotato. This study thus lays the foundation for further illustration of IbMYB3 function.

Regulatory Mechanism of Salicylic Acid on Seed Germination Under Salt Stress in Kale
Cao Dongdong,Chen Shanyu,Qin Yebo,Wu Huaping,Ruan Guanhai,Huang Yutao
Chin Bull Bot. 2020, 55(1):  49-61.  doi:10.11983/CBB19047
Abstract ( 144 )   HTML ( 0 )   PDF (2910KB) ( 106 )   Save
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Salinity is a major abiotic constraint affecting seed germination and plant growth. The effects of salt stress and salicylic acid (SA) and 2-aminoindan-2-phosphonic acid (SA synthesis inhibitor, AIP) on seed germination were investigated using kale (Brassica oleracea var. acephala) variety ‘Nagoya’. The results showed that the seed vigor of kale was significant decreased under 150 or 200 mmol·L -1 NaCl treatment by significantly reducing seed water uptake rate, seed vigor, and seedling quality, inhibited phenylalanine ammonia and lyase activity, decreasing endogenous SA content, and increasing hydrogen peroxide (H2O2) and superoxide anion (O2 -.) contents. SA treatment effectively alleviated the inhibition of salt stress on seed germination, promoted endogenous SA anabolism, increased seed water absorption rate and seed vigor, promoted K + and Mg 2+ absorption, and reduced the Na + content. Furthermore, SA treatment significantly increased superoxide dismutase and peroxidase activities and inhibited H2O2 and O2 -.accumulation. By contrast, AIP treatment significantly inhibited the germination process under salt stress, likely due to the reduced level of endogenous SA. This study demonstrates that exogenous SA enhances salt tolerance in kale seeds primarily through increasing the protective enzyme activities, reducing reactive oxygen species accumulation, and maintaining ion homeostasis during seed germination.

TECHNIQUES AND METHODS
Pull-down and Co-immunoprecipitation Assays of Interacting Proteins in Plants
Xu Chongyi
Chin Bull Bot. 2020, 55(1):  62-68.  doi:10.11983/CBB19143
Abstract ( 1692 )   HTML ( 1 )   PDF (3878KB) ( 1973 )   Save
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Protein-protein interactions play a key role in cellular signaling, involved in various biological processes. Studies on these interactions are therefore crucial toward understanding the regulatory networks of cellular signaling. It is a standard practice that the protein-protein interactions identified by the yeast two-hybrid system should be independently confirmed by in vitro and in vivo approaches. Pull-down and co-immunoprecipitation (Co-IP) are routine approaches to detect protein-protein interactions. Pull-down assay is used to detect direct or physical interactions between proteins in vitro. In plant biology studies, one of the most convenient methods to detect protein-protein interactions is the transient expression of the target proteins in Nicotiana benthamiana leaves followed by the Co-IP assay. In this paper, we describe the principles and protocols for the GST tag-based pull-down assay and the Co-IP assay of proteins transiently expressed in N. benthamiana leaves, providing a reference for detecting plant protein-protein interactions.

Luciferase Complementation Assay for Detecting Protein Interactions
Zhao Yan,Zhou Jianmin
Chin Bull Bot. 2020, 55(1):  69-75.  doi:10.11983/CBB19229
Abstract ( 1118 )   HTML ( 3 )   PDF (1554KB) ( 1347 )   Save
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Protein-Protein interactions play important roles in various eukaryotic biological processes. Compared to other techniques measuring protein-protein interactions in plants, the Luciferase Complementation Assay (LCA), based on Agrobacterium-mediated transient expression in Nicotiana benthamiana, is a simple, sensitive, reliable, highly quantitative and low background method that can be easily scaled up for high-throughput interactome studies. Here, we describe a protocol that includes two alternative data collection methods to qualitative and quantitative analyse luminescence or luminous intensity to detect protein-protein interactions in plant cells.

Protocols for Analyzing Plant Phospho-proteins
Zhu Dan,Cao Hanwei,Li Yuan,Ren Dongtao
Chin Bull Bot. 2020, 55(1):  76-82.  doi:10.11983/CBB19208
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Protein phosphorylation is one of the important protein posttranslational modifications that is involved in the regulation of most cellular processes in plants. Protein kinases catalyze the phosphorylation by transferring the phosphate group in ATP to the substrate proteins. The phosphate is usually covalently linked to the hydroxyl group of specific amino acid residues in the substrates by an ester bond. The mostly studied phosphorylation sites are serine, threonine, and tyrosine residues. Here, we present protocols and related tips for the in vitro and in vivo protein phosphorylation assays.

Detection of SUMOylation in Plants
Qu Gaoping,Jin Jingbo
Chin Bull Bot. 2020, 55(1):  83-89.  doi:10.11983/CBB19213
Abstract ( 322 )   HTML ( 3 )   PDF (1533KB) ( 408 )   Save
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SUMOylation, a post-translational modification, is essential for plant normal growth and development. To date, thousands of possible SUMO substrates have been identified, but due to the relatively low SUMOylation level, biological significance of the SUMOylation remains largely unknown. Here, we summarized the SUMOylation detection methods, including in vitro and in vivo SUMOylation assays, which help to understand the functions of SUMOylation in plants.

Optimization of Cell Suspension Culture Conditions of Achyranthes bidentata
Li Ping,Dong Yahui,Li Chenglong,He Yulong,Li Mingjun
Chin Bull Bot. 2020, 55(1):  90-95.  doi:10.11983/CBB19168
Abstract ( 121 )   HTML ( 1 )   PDF (854KB) ( 76 )   Save
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To optimize the suspension culture conditions of Achyranthes bidentata cells, effect of different factors like inoculum concentrations, subculture cycles, pH, light and Cu 2+ on cell growth and polysaccharides contents was studied. The results showed that, cells grew well with a good ability for polysaccharides synthesis under 50 g·L -1 inoculum concentration, 14 d subculture cycle, pH5-6 and light. In the presence of 50 μmol·L -1 Cu 2+, the cell dry weight could be the maximum with 44.63 g·L -1, and polysaccharides contents reached the peak at 4.02 mg·g -1.

SPECIAL TOPICS
Research Advances on Lectin Receptor-like Kinases in Plants
Wang Menglong,Peng Xiaoqun,Chen Zhufeng,Tang Xiaoyan
Chin Bull Bot. 2020, 55(1):  96-105.  doi:10.11983/CBB19130
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Plant growth and development are affected by various environmental factors. In response to various environmental changes, plants have evolved a series of signal recognition and transduction proteins, such as the plasma membrane-localized receptor-like kinases (RLKs), to cope with the environmental conditions. The lectin receptor-like kinases (LecRLKs) are a subfamily of RLKs that contain three structural domains: the extracellular lectin domain, transmembrane domain, and the intracellular kinase domain. Based on the structural difference of the extracellular lectin domain, LecRLKs are classified into three subclasses: L-, G-, and C-type. Recent studies have shown that LecRLKs play a vital role in plant development and biotic/abiotic stress responses. In this review, we discribe the research history, structural features and classification, and biological functions of LecRLKs, and emphasize on the functions of LecRLKs in plants in response to biotic/abiotic stresses and in regulating development. This review provides a view for future functional study on LecRLKs and crop improvement by elaborating different types and functions of LecRLKs.

Recent Advances in Phenolic Metabolites in Pecan
Jia Xiaodong,Xu Mengyang,Mo Zhenghai,Xuan Jiping,Zhai Min,Guo Zhongren
Chin Bull Bot. 2020, 55(1):  106-119.  doi:10.11983/CBB19008
Abstract ( 165 )   HTML ( 1 )   PDF (1199KB) ( 95 )   Save
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Pecan (Carya illinoensis) is enriched in phenolic metabolites, which have significant antioxidant activity. In recent years, the pecan industry has developed rapidly in China, with drastic increase of planting area. It is of great significance to clarify the composition and health benefits of phenolic metabolites in pecan for promoting the healthy development of the industry. So far, a total of 67 phenolic metabolites have been detected from pecan, including 36 tannins, 22 flavonoids, and 9 phenolic acids. And 34 phenolic metabolite aglycones have been detected after hydrolysis. This paper summarizes the recent progresses in the composition, extraction method, content, pharmacological activity, and relationship with postharvest of phenolic metabolites in pecan. These data are of great significance to further study the relationship between the phenolic metabolites and their physiological activities, and explore the health benefits of the phenolic metabolites in pecan.

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