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Journal of Integrative Agriculture 2023年第11期22卷 3284-3295页

作者： SU He-nan YUAN Yu-xiang YANG Shuang-juan WEI Xiao-chun ZHAO Yan-yan WANG Zhi-yong QIN Liu-yue YANG Zhi-yuan NIU Liu-jing LI Lin ZHANG Xiao-wei Institute of Horticulture Henan Academy of Agricultural SciencesZhengzhou 450002P.R.China School of Materials Science and Engineering Central South UniversityChangsha 410083P.R.China

Chinese cabbage is an economically important Brassica vegetable worldwide, and clubroot, which is caused by the soilborne protist plant pathogen Plasmodiophora brassicae is regarded as a destructive disease to Brassica crops. Previous studies on the gene transcripts related to Chinese cabbage resistance to clubroot mainly employed RNA-seq technology,although it cannot provide accurate transcript assembly and structural information. In this study, PacBio RS II SMRT sequencing was used to generate full-length transcriptomes of mixed roots at 0, 2, 5, 8, 13, and 22 days after P. brassicae infection in the clubroot-resistant line DH40R. Overall, 39 376 high-quality isoforms and 26 270 open reading frames(ORFs) were identified from the SMRT sequencing data. Additionally, 426 annotated long noncoding RNAs(lncRNAs),56 transcription factor(TF) families, 1 883 genes with poly(A) sites and 1 691 alternative splicing(AS) events were identified. Furthermore, 1 201 of the genes had at least one AS event in DH40R. A comparison with RNA-seq data revealed six differentially expressed AS genes(one for disease resistance and five for defensive response) that are potentially involved in P. brassicae resistance. The results of this study provide valuable resources for basic research on clubroot resistance in Chinese cabbage.

关键词： Chinese cabbage clubroot full-length transcriptome SMRT sequencing alternative splicing

Effects of different rotation patterns on the occurrence of clubroot disease and diversity of rhizosphere microbes

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Journal of Integrative Agriculture 2020年第9期19卷 2265-2273页

作者： YANG Xiao-xiang HUANG Xiao-qin WU Wen-xian XIANG Yun-jia DU Lei ZHANG Lei LIU Yong Institute of Plant Protection Sichuan Academy of Agricultural SciencesChengdu 610066P.R.China Key Laboratory of Integrated Pest Management on Crops in Southwest Ministry of Agriculture and Rural AffairsChengdu 610066P.R.China

clubroot disease, caused by Plasmodiophora brassicae, is one of the most destructive soil-borne diseases in cruciferous crops worldwide. New strategies are urgently needed to control this disease, as no effective disease-resistant varieties or chemical control agents exist. Previously, we found that the incidence rate and disease index of clubroot in oilseed rape decreased by 50 and 40%, respectively, when oilseed rape was planted after soybean. In order to understand how different rotation patterns affect the occurrence of clubroot in oilseed rape, high-throughput sequencing was used to analyze the rhizosphere microbial community of oilseed rape planted after leguminous (soybean, clover), gramineous (rice, maize) and cruciferous (oilseed rape, Chinese cabbage) crops. Results showed that planting soybeans before oilseed rape significantly increased the population density of microbes that could inhibit P. brassicae (e.g., Sphingomonas, Bacillus, Streptomyces and Trichoderma). Conversely, consecutive cultivation of cruciferous crops significantly accumulated plant pathogens, including P. brassicae, Olpidium and Colletotrichum (Pclubroot damage.

关键词： oilseed rape clubroot Gramineae Leguminosae Cruciferae rhizosphere soil

clubroot resistance introgression in interspecific hybrids between Raphanus sativus and Brassica napus

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Oil Crop Science 2019年第3期4卷 139-151页

作者： Xiuzhen Wang Lingyi Zeng Li Xu Wang Chen Fan Liu Huan Yang Peng Chi Li Ren Ruibin Yan Guangyuan Lu Xiaoping Fang Key Laboratory of Biology and Genetic Improvement of Oil Crops Ministry of Agriculture Oil Crops Research InstituteChinese Academy of Agricultural Sciences

clubroot is a prevailing soil-borne disease affecting rapeseed production ***,few clubroot resistant rapeseed accessions were available for *** and introgression of new clubroot resistant genes from closely related species by distant hybridization is an effective *** the present study,9 radish(Raphanus sativus L.,2n=18,RR)lines resistant to Plasmodiophora brassicae pathotype 4 were used as donors to transfer clubroot resistance into a susceptible rapeseed(Brassica napus L.,2n=38,AACC)line by distant hybridization combined with embryo *** intergeneric crosses were made but only 1(411×93039)produced F1 plants both from embryo rescue and natural *** of triploid F1 hybrids(2n=28,ACR)were verified by flower color,cytological observation and molecular marker analysis,and 2 genuine F1 hybrids were *** chromosome doubling,these synthetic allohexaploid plants(2n=56,AACCRR)became partially fertile(pollen viability rate=35%)and were backcrossed with rapeseed parent to generate a BC1 population(2n=47,AACCR).Totally 178 BC1 plants were obtained,of which the majority(96.1%)were resistant to *** backcrossing progenies could be used for the breeding of new rapeseed varieties resistant to clubroot.

关键词： clubroot Plasmodiophora brassicae Brassica napus radish intergenerichybridization

Identification of Genes Associated with clubroot Resistance in Chinese Cabbage

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Journal of Northeast Agricultural University(English Edition) 2018年第4期25卷 8-21页

作者： Song Bo Xu Hai He Jiang-ming Hu Jing-feng Fan Xiao-xue Chen Long-zheng Institute of Vegetable Crops Jiangsu Academy of Agricultural Sciences Institute of Horticultura Crops Yunnan Academy of Agricultural Sciences

The transcriptome-wide gene expression was compared between susceptible and resistant Chinese cabbage cultivars to identify genes that contributed to clubroot resistance. A higher number of differentially expressed genes were detected in susceptible cultivars than in resistant cultivars. Fifty-six genes involved in cell wall modification, hormone signaling, root marphogenesis, nematodes response and cell proliferation were uniquely expressed in the resistant cultivars. Among them, 27 genes were involved in cell wall modification and hormone signaling, indicating that genes in these two types might play a vital role in the defense response to pathogen infection.

关键词： Chinese cabbage clubroot transcriptome analysis resistant gene discovery

Influence of 6-BA on Incidence Rate of Chinese Cabbage clubroot (Plasmodiophora brassicae Woron)

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Plant Diseases and Pests 2011年第5期2卷 21-24页

作者：王哲郑萍刘照斌马有会王火旭辽宁师范大学生命科学学院辽宁大连116081 临西县技工学校河北邢台054900 辽宁师范大学分析测试中心辽宁大连116029 辽宁师范大学生命科学学院辽宁省生物技术重点实验室辽宁大连116081

[Objective] The paper was to study the relationship between cytokinin and Chinese cabbage clubroot, and to explore the incidence mechanism of clubroot. [Method]The spores were extracted from the tumors of diseased plant, and their DNA was extracted for PCR test. The diagnosed spores were inoculated to the culture soils in erlenmeyer flask at seeding stage, at seed germination and 21 d after sowing, and 0.08 μmol/L 6-BA was added to the soil at germination. Spores were extracted from the root tumors developed in inoculated plants and examined by scanning electronic microscope （ SEM）, and the incidence rate of clubroot in 6-BA treatment and control was recorded. [ Result] Plasmodiophora brassicae Woron could cause the clubroot incidence of Chinese cabbage planted in erlenmeyer flasks, the incidence rate of chibroot in the treatment adding with 0.08 μmol/L 6-BA was 100%, while that in the treatment without 6-BA was 57%, and the volume of former tumor was much larger than the latter. SEM showed that the size of resting spore of P. brassicae was 1.5 -4.3μm. [ Conduslon] 6-BA applied at germination could significantly promote the formation of tumor of Chinese cabbage clubroot.

关键词： 6-BA Chinese cabbage clubroot Scanning electronic microscope PCR technology China

Resynthesis of clubroot disease resistant rapeseed(AACr Cr and Ar Ar Cr Cr) through hybridization

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Resynthesis of clubroot disease resistant rapeseed(AACr Cr a...

Breeding of a New clubroot-resistant Chinese Cabbage Variety, CCR11242

中国植物病理学会2019年学术年会

作者： Zeng Lingyi Wang Xiuzhen Xu Li Yang Huan Chen Wang Liu Fan Yan Ruibin Ren Li Chen Kunrong Fang Xiaoping Key Laboratory of Biology and Genetic Improvement of Oil Crops Ministry of Agriculture Oil Crops Research Institute Chinese Academy of Agricultural Sciences

clubroot is a prevailing soil-borne disease affecting rapeseed production ***,very few clubroot resistant rapeseed accessions were available for the breeding of new *** and introgression of new clubroot resistant genes from closely related species by distance hybridization is an effective strategy to control this *** napus L.(AACC) is an amphidiploid species that originated from a spontaneous hybridisation of *** L.(AA) and *** L.,(CC) and contains the complete diploid chromosome sets of both parental *** this research,27 resistant/susceptible *** and 3 resistant *** varieties were identified to create new resources of clubroot disease resistant *** using resistant ***(CC) as pattermal plants and resistant/susceptible ***(AA and AA) as maternal plants,***(AAC C and AACC) was artificially resynthesized through hybridization,ovary culture and chromosome *** backcrossed to maternal plants,detection of disease resistance related molecular markers and disease resistant identification,a total of 10 lines of resynthesized *** were *** study will provide new sources of clubroot resistance materials for the breeding of new rapeseed varieties.

关键词： clubroot rapeseed AACrCr ArArCrCr hybridization

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Agricultural Biotechnology 2016年第5期5卷 1-3页

作者： Xuezhong XU Zhimei TIAN Hongli YANG Jingfeng HU Liqin ZHANG Mei LAN Hongju DUAN Jiangming HE Institute of Horticultural Crops Yunnan Academy of Agricultural Sciences Kunming 650205 China Yunnan Branch of the National Vegetable Improvement Center Kunming 650205 China Institute of Cash Crops Yunnan Academy of Agricultural Sciences Kunming 650205 China Agricultural Service Center of Xiangcheng Town Xiangyun 672100 China

In this study, using Japanese radish cytoplasmic sterile line ＂Half Green Cabbage＂（HGC） and clubfoot-resistant South Korean cabbage variety ＂ CR Cabbage King＂（CCK） as experimental materials, serf-incompatible DH lines were obtained by mierosporo culture. Through five gnerations of backeross, CCR11239, a clubroot-resistant radish cytoplasmic sterile line, was obtained. Through six generations of serf-crossing, CCR11240, the maintainer line of CCR11239, was bred. After the cross of CCK and Luchunbai 1 （83 - 1 ） and five generations of self-crossing, serf-compatible line CCRl1241 was obtained. Through crossing CCR11240 with CCRl1241, the new cabbage variety CCRl1242 was obtained. After variety comparison test, provincial regional test and production demonstration test, the new variety was registered in 2012. The average yield of CCR11242 reached 60 975 kg/hm2, which was improved by 142% compared with Luchunbai 1 （83 - 1 ）. The disease index of CCR11242 was 5.63, which was 88.13 lower than the control, indicating high resistance （HR） to clubfoot.

关键词： Chinese cabbage clubroot CCR11242 Breeding

Development of a rapid and efficient system for CR genes identification based on hairy root transformation in Brassicaceae

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Horticultural Plant Journal 2024年第4期10卷 1049-1060页

作者： Wenlin Yu Lu Yang Yuanyuan Xiang Rongde Li Xueqing Zhou Longcai Gan Xianyu Xiang Yunyun Zhang Lei Yuan Yanqing Luo Genze Li Youning Wang Yinhua Chen Peng Chen Chunyu Zhang National Key Laboratory of Crop Genetic Improvement and College of Plant Science and Technology Huazhong Agricultural UniversityWuhanHubei 430070China National Key Laboratory for Tropical Crop Breeding Sanya Nanfan Research InstituteHainan UniversitySanyaHainan 572025China National Agro-Tech Extension and Service Center Beijing 100125China Industrial Crops Research Institute Yunnan Academy of Agricultural SciencesKunmingYunnan 650221China College ofAgronomy NorthwestA&F UniversityYanglingShaanxi712100China

Many economically important crops and vegetables belonging to the cruciferous family are heavily endangered by clubroot disease caused by Plasmodiophora brassicae *** of clubroot resistant cultivars based on mapping and cloning of resistant genes is commonly regarded as the most cost-effective and efficient way to fight against this *** traditional way of R gene functional validation requires stable transformation that is both time-and *** this study,a rapid and efficient hairy-root transgenic protocol mediated by Agrobacterium rhizogenes was *** transformation positive rate was over 80%in Brassica napus showed by GUS reporter gene and this transformation only took 1/6 of the time compared with stable *** system was applicable to different *** varieties and other cruciferous crops including Brassica rapa and Brassica *** particular,two known CR genes,CRA3.7.1 and CRA8.2.4 were used respectively,as example to show that the system works well for CR gene study combined with subsequent *** infection in *** importantly,it works both in over-expression that led to disease resistance,as well as in RNAi which led to disease susceptible ***,this system can be used in batch-wise identification of CR genes,and also offered the possibility of manipulating key genes within the *** genome that could improve our knowledge on host-pathogen interaction.

关键词： Brassicaceae Agrobacterium rhizogenes Hairy root transformation clubroot Gene function

Genome-wide identification and analysis of cytokinin dehydrogenase/oxidase(CKX) family genes in Brassica oleracea *** their involvement in response to Plasmodiophora brassicae infections

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Horticultural Plant Journal 2022年第1期8卷 68-80页

作者： Mingzhao Zhu Yong Wang Shujin Lu Limei Yang Mu Zhuang Yangyong Zhang Honghao Lv Zhiyuan Fang Xilin Hou State Key Laboratory of Crop Genetics and Germplasm Enhancement College of HorticultureNanjing Agricultural UniversityNanjing 210095China Key Laboratory of Biology and Genetic Improvement of Horticultural Crops Ministry of AgricultureInstitute of Vegetables and FlowersChinese Academy of Agricultural SciencesBeijing 100081China

Cytokinins are a class of phytohormones that promote cell division and differentiation and are thought to affect plant immunity to multiple ***,a comprehensive analysis of cytokinin dehydrogenase/oxidase(CKX)family genes in cabbage has not been *** this study,a total of 36 CKX genes were identified using a genome-wide search *** analysis classified these genes into three *** were distributed unevenly across nine chromosomes in ***,and 15 of them did not contain any *** results of colinearity analysis showed that 36 CKX gene in Arabidopsis was present in several copies in the Brassica oleracea *** analysis of cisacting elements indicated that all genes possessed at least one stress or hormone responsive cis-acting element.A heatmap of CKX gene expression showed the patterns of expression of these genes in various tissues and *** genes(Bol028363,Bol031036 and Bol018140)were relatively highly expressed in all of the investigated tissues under normal conditions,showing the expression profile of housekeeping ***,the expression patterns of CKX genes in Jingfeng 1 and Xiangan 336 were quite different under the same ***,three genes(Bol020547,Bol028392 and Bol045724)were significantly down-regulated and up-regulated in the susceptible and resistant material,respectively,after inoculation,which may indicate their crucial roles in resistance to clubroot *** results provide insights for better understanding the roles of CKX genes in the ***–*** interaction.

关键词： Brassica oleracea Genome-wide Cytokinin dehydrogenase/oxidase(CKX)genes clubroot