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molecular Cloning and Characterization of a Novel Gene Involved in Fatty Acid Synthesis in Brassica napus L.

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Journal of Integrative Agriculture 2013年第6期12卷 962-970页

作者： XIAO Gang ZHANG Zhen-qian LIU Rui-yang YIN Chang-fa WU Xian-meng TAN Tai-long GUAN Chun-yun The Oil crops Research Institute/National Oil crops Improvement Center Pre-State key laboratory for germplasm innovation and Resource Utilization of crops

Based on the sequence of a novel expressed sequence tag （EST）, the full-length cDNA of 1 017 nucleotides was cloned from Brassica napus cv. Xiangyou 15 through rapid amplification of cDNA ends （RACE）. The gene was designated as Bnhol34 （HQ585980）, encoding a protein of 338 amino acids. BLAST analysis showed no high degree of sequence identity to any known gene. The calculated molecular weight of the Bnhol34 protein was 36.23 kDa, and the theoretical isoelectric point was 8.74. The Bnhol34 was also cloned from a high oleic acid mutant 854-1 through homologous cloning. There was no difference between the two Bnhol34 genes. Bnhol34 was localized in a tissue-specific manner in B. napus, and its expression level was about eight-fold greater in Xiangyou 15 seeds than in 854-1. The promoter region sequences of Bnhol34 were then isolated from Xiangyou 15 and 854-1, and a 93-bp deletion was found to occur in the Bnhol34 promoter region of 854-1. Three abscisic acid-responsive cis-elements （ABRE） were identified in the promoter region of Xiangyou 15. Real-time PCR analyses revealed that exogenous abscisic acid increased Bnhol34 expression by about four-fold in Xiangyou 15 seeds, yet did not change Bnhol34 expression in 854-1. It appeared that Bnhol34 might be abscisic acid insensitive in 854-1.

关键词： Brassca napus L. rapid amplification of cDNA ends high-efficient thermal asymmetric interlaced PCR fatty acid synthesis abscisic acid expression

SH3P2,an SH3 domain-containing protein that interacts with both Pib and AvrPib,suppresses effector-triggered,Pib-mediated immunity in rice

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molecular Plant 2022年第12期15卷 1931-1946页

作者： Yunjie Xie Yupeng Wang Xiangzhen Yu Yuelong Lin Yongsheng Zhu Jinwen Chen Hongguang Xie Qingqing Zhang Lanning Wang Yidong Wei Yanjia Xiao Qiuhua Cai Yanmei Zheng Mo Wang Huaan Xie Jianfu Zhang State key laboratory of Ecological Pest Control for Fujian and Taiwan crops Fujian Agriculture and Forestry UniversityFuzhou 350002China Rice Research Institute Fujian Academy of Agricultural SciencesFuzhou 350018China key laboratory of germplasm innovation and molecular breeding of Hybrid Rice for South China Ministry of Agriculture and AffairsFuzhouP.R.China Incubator of National key laboratory of germplasm innovation and molecular breeding between Fujian and Ministry of Sciences and Technology FuzhouChina Fuzhou Branch National Rice Improvement Center of ChinaFuzhouChina Fujian Engineering laboratory of Crop molecular breeding FuzhouChina Fujian key laboratory of Rice molecular breeding Fuzhou 350003China

Plants usually keep resistance(R)proteins in a static state under normal conditions to avoid autoimmunity and save energy for growth,but R proteins can be rapidly activated upon perceiving pathogen ***,the first cloned blast disease R gene in rice,encoding a nucleotide-binding leucine-rich repeat(NLR)protein,mediates resistance to the blast fungal(Magnaporthe oryzae)isolates carrying the avirulence gene ***,the molecular mechanisms about how Pib recognizes AvrPib and how it is inactivated and activated remain largely *** this study,through map-based cloning and CRISPR-Cas9 gene editing,we proved that Pib contributes to the blast disease resistance of rice cultivar Yunyin(YY).Furthermore,an SH3 domain-containing protein,SH3P2,was found to associate with Pib mainly at clathrin-coated vesicles in rice cells,via direct binding with the coiled-coil(CC)domain of ***,overexpression of SH3P2 in YY compromised Pib-mediated resistance to *** isolates carrying AvrPib and Pib-AvrPib recognition-induced cell ***3P2 competitively inhibits the self-association of the Pib CC domain in vitro,suggesting that binding of SH3P2 with Pib undermines its ***,SH3P2 can also interact with AvrPib and displays higher affinity to AvrPib than to Pib,which leads to dissociation of SH3P2 from Pib in the presence of *** together,our results suggest that SH3P2 functions as a“protector”to keep Pib in a static state by direct interaction during normal growth but could be triggered off by the invasion of AvrPib-carrying *** *** study reveals a new mechanism about how an NLR protein is inactivated under normal conditions but is activated upon pathogen infection.

关键词： rice blast fungus Pib-AvrPib recognition SH3 domain-containing protein protector

Preferential transport activity of Dk DTX5/MATE5 affects the formation of different astringency in persimmon

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Journal of Integrative Plant Biology 2023年第10期65卷 2304-2319页

作者： Ying Liu Xin Wu Chenfeng Sun Wenxing Chen Meng Zhang Niannian Liu Qinglin Zhang Liqing Xu Zhengrong Luo National key laboratory for germplasm innovation&Utilization of Horticultural crops Huazhong Agricultural UniversityWuhan 430070China

Proanthocyanidins(PAs) are specialized metabolites that infuence persimmon fruit *** astringent(A)-type and non-astringent(NA)-type mutants show significant variation in PA accumulation, but the infuencing mechanism remains unclear. In this study, among the six identified DTXs/MATEs proteins associated with PA accumulation, we observed that allelic variation and preferential transport by Dk DTX5/MATE5 induced variation in PA accumulation for A-type and NA-type fruit. The expression pattern of Dk DTX5/MATE5 was correlated with PA accumulation in NA-type fruit. Upregulation and downregulation of Dk DTX5/MATE5 promoted and inhibited PA accumulation, respectively, in the NA-type fruit. Interestingly, transporter assays of Xenopus laevis oocytes indicated that Dk DTX5/MATE5 preferentially transported the PA precursors catechin, epicatechin, and epicatechin gallate, resulting in their increased ratios relative to the total PAs, which was the main source of variation in PA accumulation between the A-type and NA-type. The allele lacking Ser-84 in Dk DTX5/MATE5 was identified as a dominantly expressed gene in the A-type and lost its transport function. Site-directed mutagenesis revealed that Dk DTX5/MATE5 binds to PA precursors via Ser-84. These findings clarify the association between the transporter function of Dk DTX5/MATE5 and PA variation, and can contribute to the breeding of new cultivars with improved fruit quality.

关键词： astringency DTX/MATE persimmon(Diospyros kaki Thunb.) proanthocyanidins(PAs) tonoplast transmembrane

Sdr4 dominates pre-harvest sprouting and facilitates adaptation to local climatic condition in Asian cultivated rice

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Journal of Integrative Plant Biology 2022年第6期64卷 1246-1263页

作者： Bo Zhao Hui Zhang Tianxiao Chen Ling Ding Liying Zhang Xiali Ding Jun Zhang Qian Qian Yong Xiang Shenzhen Branch Guangdong Laboratory for Lingnan Modern AgricultureGenome Analysis Laboratory of the Ministry of AgricultureAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhen 518120China shanxi key laboratory of minor Crop germplasm innovation and molecular breeding College of Life SciencesShanxi Agricultural UniversityTaigu 030801China State key laboratory of Rice Biology China National Rice Research InstituteHangzhou 310006China

Pre-harvest sprouting(PHS),which reduces grain yield and quality,is controlled by seed dormancy *** few dormancy-related genes have been cloned,the genetic basis of seed dormancy in rice(Oryza sativa L.)remains ***,we performed a genome-wide association study and linkage mapping to dissect the genetic basis of seed dormancy in *** findings suggest that Seed Dormancy4(Sdr4),a central modulator of seed dormancy,integrates the abscisic acid and gibberellic acid signaling pathways at the transcriptional *** analysis revealed that three Sdr4 alleles in rice cultivars already existed in ancestral Oryza rufipogon ***,like the semi-dwarf 1(SD1)and Rc loci,Sdr4 underwent selection during the domestication and improvement of Asian cultivated *** distribution frequency of the Sdr4-n allele in different locations in Asia is negatively associated with local annual temperature and ***,we developed functional molecular markers for Sdr4,SD1,and Rc for use in molecular *** results provide clues about the molecular basis of Sdr4-regulated seed ***,these findings provide guidance for utilizing the favorable alleles of Sdr4 and Rc to synergistically boost PHS resistance,yield,and quality in modern rice varieties.

关键词： climatic adaptation pre-harvest sprouting Rc rice SD1 Sdr4 seed dormancy

Identification of Flowering Regulatory Genes in Allopolyploid Brassica juncea

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Horticultural Plant Journal 2019年第3期5卷 109-119页

作者： Ning Zhao Chang Liu Yiqing Meng Zhongyuan Hu Mingfang Zhang Jinghua Yang laboratory of germplasm innovation and molecular breeding Institute of Vegetable Science Zhejiang University key laboratory of Horticultural Plant Growth Development & Quality Improvement Ministry of Agriculture

Brassica juncea is an allopolyploid originating from the interspecific hybridization between Brassica rapa and Brassica nigra, which is of multiple usage as a vegetable, oilseed and condiment worldwide. Both vernalization and non-vernalization under long-day photoperiod can promote floral transition in B. juncea suggesting merged flowering pathways of its ancestors and better environmental adaptability. We identified genomewide flowering regulatory genes in B. juncea, which include 84 and 79 genes from A and B sub-genomes, respectively. Ka/Ks analysis revealed a purification effect on both photoperiod and vernalization flowering regulation pathways during evolution. Expression profile of those genes during long-day and vernalization treatments suggested Bju ACO4, Bju AFT1, Bju BFT4, Bju ASOC1 and Bju ASOC4 may be the major functional copies of B. juncea flowering regulation. Further functional studies about Bju COs showed three days delayed flowering time in Bju ACO4 or Bju BCO3 silenced plants. Increased transcription of all BjuFLCs in Bju ACO4 or Bju BCO3 silenced plants suggested interactions between photoperiod and vernalization pathways governing flowering time. Our findings provided flowering regulating networks in allopolyploid B. juncea.

关键词： Brassica juncea Allopolyploidy Flower Vernalization Photoperiod

Transcript Profiling Reveals Abscisic Acid,Salicylic Acid and Jasmonic-Isoleucine Pathways Involved in High Regenerative Capacities of Immature Embryos Compared with Mature Seeds in japonica Rice

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Rice science 2018年第4期25卷 227-234页

作者： XIAO Kaizhuan MAO Xiaohui WANG Yingheng WANG Jinlan WEI Yidong CAI Qiuhua XIE Hua'an ZHANG Jianfu College of Life Sciences Fujian Normal UniversityFuzhou 350108China Rice Research Institute Fujian Academy of Agricultural SciencesFuzhou 350019China key laboratory of germplasm innovation and molecular breeding of Hybrid Rice for South China Ministry of Agriculture/Incubator of National Key Laboratory of Fujian Germplasm Innovation and Molecular Breeding Between Fujian and Ministry of Sciences&Technology/Fuzhou BranchNational Rice Improvement Center of China/Fujian Engineering Laboratory of Crop Molecular Breeding/Fujian Key Laboratory of Rice Molecular Breeding/Base of South ChinaState Key Laboratory of Hybrid Rice/National Rice Engineering LaboratoryFuzhou 350003China

Induced pluripotent cell mass plays a role in genetic transformation mediated by Agrobacterium. Mature seeds are more recalcitrant to the induction of suitable calli than immature embryos in rice, but the exact molecular mechanisms involved remain elusive. In this study, the morphological structure of calli induced from mature seeds and immature embryos were observed under a scanning electron microscope using a paraffin embedded technique. Meanwhile, a total of 2 173 up- and down-regulated genes were identified in calli induced from mature seeds and immature embryos by RNA-seq technique and furtherly confirmed by quantitative real-time PCR. The results revealed the remarkable morphological differences in calli induced from mature seeds and immature embryos, and plant hormone signal transduction and hormone biosynthesis pathways, such as abscisic acid, salicylic acid and jasmonic-isoleucine, were found to play roles in somatic embryogenesis. This study provided comprehensive gene expression sets for mature seeds and immature embryos that were served as an important platform resource for further functional studies in plant embryogenesis.

关键词： callus immature embryo mature seed japonica rice RNA sequence hormone

Determination of Heterotic Groups and Heterosis Analysis of Yield Performance in indica Rice

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Rice science 2018年第5期25卷 261-269页

作者： WANG Yingheng CAI Qiuhua XIE Hongguang WU Fangxi LIAN Ling HE Wei CHEN Liping XIE Hua'an ZHANG Jianfu College of Crop Sciences Fujian Agriculture and Forestry University Fuzhou 350002 China Rice Research Institute Fujian Academy of Agricultural Sciences Fuzhou 350019 China key laboratory of germplasm innovation and molecular breeding of Hybrid Rice for South China Ministry of Agriculture / Fuzhou Branch National Rice Improvement Center / Fujian Engineering Laboratory of Crop Molecular Breeding / Fujian Key Laboratory of Rice Molecular Breeding Fuzhou 350003 China Incubator of National key laboratory of Fujian germplasm innovation and molecular breeding Between Fujian and Ministry of Sciences & Technology Fuzhou 350003 China National Rice Engineering laboratory Fuzhou 350003 China

To compare the heterosis levels among various groups of parental lines used extensively in China, identify foundational heterotic groups in parental pools and understand the relationship between genetic distance and heterosis performance, 16 parental lines with extensive genetic variation were selected from various sub-groups, and 39 hybrid combinations were generated and evaluated in Fujian and Hainan Provinces of China. The main results were as follows：（1） The 16 parental lines can be grouped into 7 sub-groups consisting of 1 maintainer sub-group and 6 restorer sub-groups; （2） Mean grain yield of the restorer lines was higher than that of the maintainer lines, and mean yield of parental lines was higher than that of the hybrid combinations; （3） The two best heterotic patterns were II-32A × G5 and II-32A × G6, moreover, the order of restorer sub-groups according to grain yield, from the highest to lowest, was G7, G6, G5, G4, G3 and G2; High specific combining ability values were observed for combinations of II-32A × G5, II-32A × G6 and Tianfeng A × G7; （4） Hybrid combinations derived from II-32A crossed with 13 restorer lines had higher yield trait values （mid-parent heterosis, better-parent heterosis, standard heterosis over check and specific combining ability） than any other combinations; （5） Genetic distance was positively correlated with panicle number, grain length and length-to-width ratio （P 〈 0.05） and negatively correlated with grain width, grain yield, seed-setting rate, as well as mid-parent heterosis, standard heterosis over check, and specific combining ability for grain yield （P 〈 0.01）. These heterotic groups and patterns and their argonomic traits will provide useful information for future hybrid rice breeding programs.

关键词： rice heterosis heterosis group general combine ability specific combine ability genetic distance

Tobacco Rattle Virus-induced Phytoene Desaturase (PDS) Silencing in Centaurea cyanus

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Horticultural Plant Journal 2021年第2期7卷 159-166页

作者： Chengyan Deng Fan Zhang Jiaying Wang Yanfei Li He Huang Silan Dai Beijing key laboratory of Ornamental Plants germplasm innovation&molecular breeding National Engineering Research Center for FloricultureBeijing Laboratory of Urban and Rural Ecological Environment and College of Landscape ArchitectureBeijing Advanced Innovation Center for Tree Breeding by Molecular DesignBeijing Forestry UniversityBeijing 100083China

Virus-induced gene silencing(VIGS)is a genetic tool used to assess gene *** rattle virus(TRV)is a VIGS vector commonly used to induce endogenous gene silencing in ***,there is no VIGS system established for Centaurea *** evaluated the effectiveness of a TRV-based VIGS system using phytoene desaturase(PDS)as a reporter gene in Centaurea *** methods including pressure-,vacuum-and apical meristem-infiltrationwere tested to infect *** *** leaveswere only obtained using apicalmeristem-infiltration after a 14 d *** CcPDS transcripts in photobleached leaves were significantly reduced compared with that in green leaves treated with empty *** *** cultivars were tested to detect their VIGS responses,and‘Dwarf Tom Pouce Blue’was the most *** agro-infiltration condition was optimized by screening for the optimal seedling stage as well as the optimum Agrobacterium density for efficient *** with four true leaves and infiltration with an Agrobacterium density of OD_(600)0.5 were optimal conditions to obtain more photobleached leaves and more intense photobleached *** results demonstrated the feasibility of TRV-based VIGS for functional analysis of genes in ***.

关键词： Centaurea cyanus VIGS tobacco rattle virus PDS seedling photobleached

Development and application of microsatellite markers within transcription factors in flare tree peony(Paeonia rockii)based on next-generation and single-molecule long-read RNA-seq

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Journal of Integrative Agriculture 2021年第7期20卷 1832-1848页

作者： LIU Na CHENG Fang-yun GUO Xin ZHONG Yuan Beijing Advanced innovation Center for Tree breeding by molecular Design/Beijing key laboratory of Ornamental Plants germplasm innovation&molecular breeding/National Engineering Research Center for Floriculture/key laboratory of Genetics and breeding in Forest Trees and Ornamental Plants of Ministry of Education/Peony International Institute School of Landscape ArchitectureBeijing Forestry UniversityBeijing 100083P.R.China

Tree peonies native to China are a precious crop with ornamental,medicinal and edible oil properties,of which flare tree peony(Paeonia rockii)is one of the most significant germplasms in *** development and application of expressed sequence tag-simple sequence repeat(EST-SSR)markers are very valuable for genetic and breeding applications,but EST-SSR resources for the genus Paeonia are still *** this study,we first reported the development of SSRs within transcription factors(TFs)in *** based on next-generation sequencing(NGS)and single-molecule long-read sequencing(SMLRS).A total of 166 EST-SSRs containing six nucleotide repeat types were identified from 959 candidate TFs associated with yield,with an average of one SSR per 5.83 *** total,102(61.45%)pairs of primers produced amplification products in the two RNA-seq *** them,58(56.86%)pairs of primers from 18 gene families(AP2,b HLH,HSF,etc.)were identified to be polymorphic both in the parents of a linkage mapping population and in eight randomly selected accessions of ***,the 58 EST-SSRs indicated a high level of informativeness with PIC values ranging from 0.32 to 0.91(mean 0.70)after assessment in 37 tree peony *** studies indicated that the amplification ratio of the 58 pairs of primers ranged from 89.66 to 100%across seven species of *** addition,a genetic relationship study was performed in 62 *** analysis using the neighbour-joining(NJ)tree demonstrated that major clusters corresponded to the known pedigree *** together,these newly developed EST-SSRs have a potential use in the conservation of tree peony germplasm and marker-assisted selection(MAS)breeding.

关键词： flare tree peony(Paeonia rockii) EST-SSR markers polymorphism genetic diversity transferability genetic relationships