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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 56 Issue 4
01 July 2021
  
EXPERIMENTAL COMMUNICATIONS
Effects of Nitrogen Application on Nitrogen Fixation Capacity and GmLbs Expression in Soybean
Mengke Du, Wenting Lian, Xiao Zhang, Xinxin Li
Chinese Bulletin of Botany. 2021, 56(4):  391-403.  doi:10.11983/CBB21037
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The nitrogen fixation efficiency of symbiotic nodules is tightly regulated by external nitrogen (N). In addition to nitrogenase activity, the leghemoglobin (Lb) amount is vital index contributing to N2 fixation. To determine the effect of environmental N level on biological nitrogen fixation, soybean plants were inoculated with rhizobia for 30 d under low N condition (0.53 mmol·L-1), then transplanted to hydroponic culture solution with relatively higher N concentrations, including 5.3, 10, 20, 30, and 40 mmol·L-1 for 7 d, respectively. Lb concentration, nitrogenase activity and bacteroid development status were measured. Nodule displayed changed color from red to green with increasing N concentrations. Consistently, the red Lb concentration gradually declined with increased green Lb concentration. Moreover, the nitrogenase activity, infected cell number and area in nodules were all significantly decreased, suggesting that changes in Lb forms caused by excess N are closely associated with nitrogen fixation capacity. Bioinformatics and public expression profile data displayed that four symbiosis-associated Lbs, including GmLb1, GmLb2, GmLb3, and GmLb4 are the major Lb genes in soybean nodulation. These four GmLbs belong to the same clade of phylogenetic tree. Further analysis on the transcripts of GmLb1-4 in response to N showed that the expression levels of GmLb1-4 were all significantly inhibited by high N. This study services as a reference for future studies in understanding the underlying mechanisms of N-triggered nodule senescence as well as BNF application in agriculture.

AtMYB77 Involves in Lateral Root Development via Regulating Nitric Oxide Biosynthesis under Drought Stress in Arabidopsis thaliana
Yongmei Che, Yanjun Sun, Songchong Lu, Lixia Hou, Xinxin Fan, Xin Liu
Chinese Bulletin of Botany. 2021, 56(4):  404-413.  doi:10.11983/CBB20207
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Both transcription factor MYB77 and signal molecule nitric oxide (NO) are important regulators of lateral root development. However, our understanding about the role of MYB77 and NO in the regulation of lateral root formation in plants remains elusive. This study investigated the roles and interrelation of MYB77 and NO in regulating lateral root formation under drought stress by using wild type Arabidopsis, AtMYB77 deletion mutant Atmyb77-1 and overexpression lines AtOE77-1 and AtOE77-3. The results showed that the expression of AtMYB77 was induced by drought stress. When subjected to drought stress treatment, the Atmyb77-1 mutant showed down-regulation of CYCA2;1 and CDKA;1, two genes that are related with lateral root development. Meanwhile, the number and length of lateral roots in the Atmyb77-1 mutant were significantly lower than those in wild type, while AtOE77-1 and AtOE77-3 lines displayed more and longer lateral roots. These results indicated that AtMYB77 was involved in the regulation of lateral root development under drought stress. We also showed that drought stress could increase the NO content, as well as the nitric oxide synthase (NOS) and nitrate reductase (NR) enzymes activity and gene expression in roots of Arabidopsis. Such increase in NO content, NOS and NR activities as well as related gene transcript levels were attenuated by deletion of AtMYB77 but enhanced by AtMYB77 overexpression. Exogenous NO donor sodium nitroprusside (SNP) alleviated the inhibitive effects of AtMYB77 deletion on the expressions of CYCA2;1 and CDKA;1 as well as the lateral root formation, while NO sca-vengers or synthesis inhibitors attenuate the promoting effect of AtMYB77 overexpression on lateral root growth. Taken together, these results demonstrate that AtMYB77 participates in drought-induced lateral root growth by promoting NO synthesis.

Heterologous Overexpression of Desiccation-tolerance Moss ScABI3 Gene Changes Stomatal Phenotype and Improves Drought Resistance in Transgenic Arabidopsis
Yigong Zhang, Yi Zhang, Ayibaiheremu Mutailifu, Daoyuan Zhang
Chinese Bulletin of Botany. 2021, 56(4):  414-421.  doi:10.11983/CBB20212
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ABI3 is a key transcriptional factor of the ABA signaling pathway, which is involved in seed dormancy, plastid development, and desiccation tolerance of bryophytes, and plays an important role in the stress tolerance of plant. This study obtained three independent pCAMBIA1301-ScABI3 transgenic Arabidopsis homozygous lines. The results showed that transgenic plants increased leaf stomata diameter, reduced the number of stomata in unit area, and improved plant water use efficiency; transgenic plants survival rate was significantly higher than wild type (WT) after 14 days of drought treatment; the water loss rate of transgenic leaves was significantly lower than that of WT. Further research has found that ScABI3 transgenic plants had higher drought resistance characteristics by improving reactive oxygen species (ROS) scavenging ability. These results may be contributed to the development and utilization of genetic resources of desert plants, also provide the theoretical and practical base for molecular breeding.

Clustering Analysis of Volatile Components from the Tea Plants Infested by Tea Aphid (Toxoptera aurantii)
Tingzhe Sun, Zehua Qi, Kexin Liang, Qin Li, Yuchun Rao, Dan Mu
Chinese Bulletin of Botany. 2021, 56(4):  422-432.  doi:10.11983/CBB21031
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Shuchazao tea (Camellia sinensis cv. ‘shuchazao’) is a newly cultivated tea species in southwest Anhui, China. To investigate the volatile release profiles of tea aphid (Toxoptera aurantii) infested tea shoots, gas chromatography-mass spectrometry (GC-MS) analysis was used to compare the volatile compositions and relative contents in healthy and infested tea shoots. 16 volatile organic compounds (VOCs) were detected in healthy tea shoots with fewer relative contents, whereas more volatiles (24 VOCs) were released with increasing relative contents in tea plants induced by tea aphid feeding. Unsupervised clustering analysis based on the significantly different volatiles indicated that the volatile profiles in healthy and tea aphid infested tea shoots apparently showed clustering characteristics. A model was created by supervised partial least square discrimination analysis (PLS-DA) for volatiles with statistical significance. By experimental verification, the model could clearly discriminate healthy tea shoots from aphid infested ones (R2X=0.903, R2Y=0.875). By calculating the variable importance for the projection (VIP), seven important volatiles (α-pinene, longifolene-(V4), benaldehyde, E-5,9-undecadien-2-one,6,10-dimethyl-, 3-hexenal, camphor and decanal) were identified which collectively contribute to discrimination between healthy and infested tea shoots. The current work has preliminarily demonstrated the changing patterns in tea plant volatiles after tea aphid infestation to provide novel theoretical guidance for tea aphid management.

Identification and Development of Polymorphic Genic-SSRs in Tamarix ramosissima in Alxa Region Based on Transcriptome
Yanan Zhang, Lei Huang, Jiabin Li, Lei Zhang, Zhenhua Dang
Chinese Bulletin of Botany. 2021, 56(4):  433-442.  doi:10.11983/CBB20185
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Simple sequence repeats located in gene transcribed regions (Genic-SSR) can play important roles in plant adaptation to environmental changes. In this study, the transcriptomes of Tamarix ramosissima from five different locations in Alax were sequenced, assembled, and compared. By using CandiSSR software, a total of 1 185 polymorphic Genic-SSRs representing 157 motif types were identified in 1 123 transcripts. Among them, the trinucleotide repeats (596, 50.30%) were the most abundant, followed by dinucleotide repeats (486, 41.01%). Location analysis showed that 411, 239, and 163 Genic-SSRs were located in CDSs, 5′UTRs, and 3′UTRs of the relevant transcripts, respectively; 78.47% of the trinucleotide SSRs were located in CDSs, and 94.07% of the dinucleotide SSRs were located in UTRs. Among SSRs distributed in CDSs, AGC/GCT, AGG/CCT, AAG/CTT, CCG/CGG, and ATC/GAT were relatively abundant, accounting for 64.48% of all the Genic-SSRs; AG/CT and AT/AT were the most abundant repeat types in UTRs, which together account for 55.22% of all the Genic-SSRs in UTRs. Functional annotation showed that polymorphic Genic-SSRs containing genes enriched in a wide range of GO terms and KEGG pathways that highly related to stress response in T. ramosissima. Of the 15 randomly selected Genic-SSRs, 14 were successfully amplified by using polymerase chain reaction technology and 64 alleles were found in these SSR loci. Genetic polymorphism estimation showed that the mean of expected and observed heterozygosity (He, Ho), polymorphism information content (PIC) of these SSRs were 0.553, 0.421, and 0.493, respectively, demonstrating the feasibility of developing SSR markers by RNA-seq.

TECHNIQUES AND METHODS
Callus Induction and Somatic Embryogenesis in Anther Culture of Paeonia lactiflora
Yanmin Li, Hui Jiang, Zhenzhu Fu, Jing Zhang, Xin Yuan, Huijuan Wang, Jie Gao, Xiaoyu Dong, Limin Wang, Hechen Zhang
Chinese Bulletin of Botany. 2021, 56(4):  443-450.  doi:10.11983/CBB20195
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The anthers of Paeonia lactiflora cv. ‘Fenyunu’ were used as explants to study the effects of different concentrations of 2,4-D on callus induction, somatic embryogenesis and plant regeneration. The cell composition of callus and the development process of somatic embryos were observed with cytohistological method, and the ploidy of regenerated plants was identified using root tip squash method. The results showed that the suitable medium for callus induction of P. lactiflora anther was MS+1 mg·L-12,4-D+1 mg·L-1NAA+0.1 mg·L-1KT+30 g·L-1sucrose+6.5 g·L-1agar, and the callus induction rate was 14.7%. The callus was transferred to somatic embryo induction medium and underwent stages of spherical embryo, heart-shaped embryo, torpedo embryo and cotyledon embryo, and the somatic embryo induction rate was 52.1%. Genuine leaves germinated in seedling medium and complete plants were obtained, and the seedling rate was 47.1%. Haploid and diploid plants were identified using root tip squash method. The study preliminarily established a culture system to implement plant regeneration through somatic embryogenesis, which also provided reference protocol for anther culture of other varieties of Paeonia. Regenerated plants are important materials for genetic research and haploid breeding of P. lactiflora.

Callus Induction and Plant Regeneration of Cerasus serrulata var. lannesiana cv. ‘Grandiflora’
Qian Luo, Yansha Zhang, Jing Ou
Chinese Bulletin of Botany. 2021, 56(4):  451-461.  doi:10.11983/CBB20205
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In order to establish the regeneration system of Cerasus serrulata var. lannesiana cv. ‘Grandiflora’, the effects of different explants and plant hormone combinations on callus induction, adventitious bud differentiation, proliferation and rooting were studied using perennial mother plant leaflets, annual grafted seedling leaflets, axillary bud induction leaflets and proliferative first-generation leaflets as explants. The results showed that callus could be induced from all four explants, and adventitious buds could be derived from all explants except the leaflets of perennial mother plants. The higher the degree of explants’ juvenility, the greater the success of subsequent culture, with the best explants were the proliferative first generation leaflets. The best medium for callus induction was MS+0.5 mg·L-1 6-BA+1.0 mg·L-1 2,4-D, and the induction rate was 96.22%. The optimal medium for differentiation was MS+1.0 mg·L-1 6-BA+0.1 mg·L-1 2,4-D+0.1 mg·L-1 TDZ, and the differentiation rate was 78.14%. The optimal medium for proliferation was MS+1.0 mg·L-1 6-BA, and the proliferation coefficient reached 7.85. The optimal medium for rooting was 1/2MS without any hormone, and the regenerated plants with 100% rooting rate were obtained. The regenerated plants from different explants grew very differently, and those induced from first generation of proliferative leaflets had the best growth.

SPECIAL TOPICS
Research Progress of CURT1 on Regulating Thylakoid Membrane Curvature
Qiuxin Li, Wei Chi, Daili Ji
Chinese Bulletin of Botany. 2021, 56(4):  462-469.  doi:10.11983/CBB20194
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The grana in chloroplast of higher plants is a structure composed of many thylakoid discs stacked together. The formation of the grana facilitates the distribution of the photosynthetic protein complex in different positions in thylakoids, that is, it has lateral heterogeneity and can effectively carry out photosynthesis. The key step to promote the formation of grana is to bend the thylakoid membrane. CURVATURE THYLAKOID 1 (CURT1) protein has been found to be the key factor leading to membrane bending. In this review, the recent research progresses of CURT1 protein in Arabidopsis thaliana and Cyanobacteria were summarized, and the prospect of CURT1 protein research in the future was put forward.

Secondary Metabolites of Syringa and the Linking with Phylogenetic Evolution and Geographical Distributions
Zhaoyu Zhang, Qingyun Wang, Lei Shi, Wengang Yu, Yongqing Zhang, Hongxia Cui
Chinese Bulletin of Botany. 2021, 56(4):  470-479.  doi:10.11983/CBB20178
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Secondary metabolites of plants induced by environmental factors are highly variable, but the given metabolic pathways may have some phylogenetic implications. Due to the difficulty in complete and systematic collections in certain plant groups, the research on the correlation between secondary metabolites and phylogeny is limited. Based on the published papers, 377 secondary metabolites in the roots, stems, leaves and flowers of Syringa were collected, which mainly derived from the mevalonic acid pathway, deoxyxylulose-5-P pathway and shikimic acid pathway. After superimposing phylogenetic background, we found that dominance of a given type of secondary metabolites was high for the firstly diverged series, and the dominance declined for subsequently diverged series with the increase of chemical diversity. Phenylpropanoids and iridoids/secoiridoids were phylogenetically conserved. After superimposing geographical distributions, we found that some local species which were lately diverged had more diverse secondary metabolites compared with widespread species firstly diverged. The high proportion of lignans was highly related to the environmental pressure. This review provided a new clue for the systematic study on the variation pattern of chemical diversity in the taxa within genus in the light of evolution.

Nucleo-cytoplasmic Transport and Transport Receptors in Plant Disease Resistance Defense Response
Tiantian Shi, Ying Gao, Huan Wang, Jun Liu
Chinese Bulletin of Botany. 2021, 56(4):  480-487.  doi:10.11983/CBB21034
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Plant pathogens pose a constant and major threat to global food production, so understanding plant’s defense mechanism against pathogen and pathogen’s infection mechanism against host crops and their molecular mechanisms will be helpful to design protection strategies for durable resistance of plant. Until now, a growing number of studies have shown that some disease resistant proteins need to be transferred to the nucleus to initiate an immune response. Nucleocytoplasmic transport receptors are essential “carrier” for nuclear transport. Therefore, nucleocytoplasmic transport and receptors play important role in disease resistance. Based on the introduction of plant disease defense response mechanism, this paper focuses on research progress of nucleocytoplasmic transport and nucleocytoplasmic transport receptors in disease resistance and proposes a prospect.

Protein Phosphorylation and Its Regulatory Roles in Seed Dormancy and Germination
Xiaoting Zhao, Kaitao Mao, Jiahui Xu, Chuan Zheng, Xiaofeng Luo, Kai Shu
Chinese Bulletin of Botany. 2021, 56(4):  488-499.  doi:10.11983/CBB21011
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Seed dormancy and germination are two distinct but closely related physiological processes, which are also key stages during plant life-cycle and have great significance to agricultural production, plant species reproduction, and geographical distribution. These processes are precisely regulated by interactions between different endogenous phytohormones and environmental signals. A large number of studies have shown that protein phosphorylation, plays an important role in regulating seed dormancy and germination, as well as plant response to stresses. This review paper briefly introduces the procedures and functions of protein phosphorylation and dephosphorylation modification, and summarizes the regulatory roles of protein phosphorylation modification in seed dormancy and germination. Finally, some future research directions are prospected.

Multispectral Imaging and Its Applications in Plant Science Research
Zhongsi Wang, Yaping Jia, Jin Zhang, Ruohan Wang
Chinese Bulletin of Botany. 2021, 56(4):  500-508.  doi:10.11983/CBB21002
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Multispectral imaging (MSI) is an emerging technology designed for advanced imaging detection, which combines the information of spectroscopy and imaging to conduct qualitative and quantitative analysis of plant phenotypes including structural, physiological and biochemical characteristics. Recently, MSI shows a strong capability to capture detailed spectral information in combination with the applications of mathematical modeling and analysis, and displays a strong potential in the field of plant research. Here we introduce the principle of MSI technology and summarize the main applications of this technology in various aspects of plant research, which includes detection of plant damage and disease, identification of plant metabolites and characterizing plant physiological status. We alse prospect the frontier development of MSI in plant research.

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