BnKIX8调控甘蓝型油菜器官大小与菌核病抗性的初步研究

doi:10.3724/SP.J.1006.2026.65014

作物学报 ›› 2026, Vol. 52 ›› Issue (11): 3334-3351.doi: 10.3724/SP.J.1006.2026.65014

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

BnKIX8调控甘蓝型油菜器官大小与菌核病抗性的初步研究

罗丽娅,马伍慧,杨龙,祝培,李艳,许本波,张学昆,徐劲松*

长江大学农学院 / 农业农村部长江中游作物绿色高效生产重点实验室(部省共建) / 涝渍灾害与湿地农业湖北省重点实验室, 湖北荆州434025

收稿日期:2026-03-02 修回日期:2026-07-15 接受日期:2026-07-15 出版日期:2026-11-12 网络出版日期:2026-07-23
通讯作者: 徐劲松, E-mail: xujinsong@yangtzeu.edu.cn
基金资助:
本研究由农业生物育种重大项目(2023ZD04042)资助。

Preliminary study on the regulation of organ size and Sclerotinia sclerotiorum resistance by BnKIX8 in rapeseed (Brassica napus L.)

Luo Li-Ya,Ma Wu-Hui,Yang Long,Zhu Pei,Li Yan,Xu Ben-Bo,Zhang Xue-Kun,Xu Jin-Song*

College of Agronomy, Yangtze University / Key Laboratory of Green and Efficient Crop Production in the Middle Reaches of Yangtze River, Ministry of Agriculture and Rural Affairs / Hubei Key Laboratory of Waterlogging Disaster and Agricultural Use of Wetland, Jingzhou 434025, Hubei, China

Received:2026-03-02 Revised:2026-07-15 Accepted:2026-07-15 Published:2026-11-12 Published online:2026-07-23
Supported by:
This study was supported by the Major Projects of Agricultural Biology Breeding of China (2023ZD04042).

摘要/Abstract

摘要：

生产中高产潜力与病害抗性常难以协同改良，探究甘蓝型油菜BnKIX8同源拷贝在生长发育和菌核病抗性中的功能及分子机制，对单株产量提升与抗病性协同改良具有重要意义。本研究以拟南芥AtKIX8为参考基因，鉴定了甘蓝型油菜中BnKIX8同源拷贝并分析其表达特征，利用CRISPR-Cas9技术创制BnKIX8s的功能缺失突变体，通过比较纯合突变体的农艺性状和菌核病抗性变化明确其功能，并结合转录组初步探究其分子机制。结果表明：(1) 甘蓝型油菜中共鉴定到4个BnKIX8同源拷贝，BnKIX8.A7和BnKIX8.C7在18个采样点均有表达，且在角果和种子中高表达，然而BnKIX8.A1和BnKIX8.C1未见有表达量。(2) CRISPR-Cas9基因编辑成功获得无T-DNA的纯合bnkix8.a7、bnkix8.c7单突变体和bnkix8.a7bnkix8.c7双突变体；表型分析表明，与WT相比，bnkix8s突变体的花朵大小、角果长度、种子大小、千粒重显著增加，种子发芽指数和活力指数显著提高；双突变体的表型呈加性效应，BnKIX8.A7和BnKIX8.C7在油菜生长发育调控中功能冗余。(3) 菌核病抗性鉴定显示，接种后36 h和48 h，bnkix8s突变体的病斑面积均比WT显著增大，表明BnKIX8.A7和BnKIX8.C7正调控菌核病抗性。(4) 转录组KEGG富集分析结果表明，在正常生长状态下，BnKIX8.A7和BnKIX8.C7可能通过影响IAA和JA信号转导通路参与“生长-防御权衡”；接种核盘菌后24 h，BnKIX8.A7和BnKIX8.C7可能通过影响α-亚麻酸代谢和硫胺素代谢通路参与菌核病抗性反应。本研究为后续深入解析BnKIX8s的功能提供了候选通路和基因资源，为油菜高产抗病育种提供了理论依据。

关键词: 甘蓝型油菜, BnKIX8, CRISPR-Cas9, 核盘菌, 转录组

Abstract: In crop production, high-yield potential and disease resistance are often difficult to synergistically improve. Exploring the functions and molecular mechanisms of BnKIX8 homologous copies in the growth, development, and Sclerotinia sclerotiorum resistance of Brassica napus is therefore of great significance for the coordinated improvement of single-plant yield and disease resistance. In this study, Arabidopsis thaliana AtKIX8 was used as the reference gene to identify BnKIX8 homologous copies in B. napus, and their expression patterns were analyzed. CRISPR-Cas9 technology was used to generate loss-of-function mutants of BnKIX8s. Their functions were clarified by comparing agronomic traits and S. sclerotiorum resistance between homozygous mutants and wild-type plants, and the underlying molecular mechanisms were preliminarily explored through transcriptome analysis. The results were as follows: (1) Four homologous copies of BnKIX8 were identified in B. napus. BnKIX8.A7 and BnKIX8.C7 were expressed at all 18 sampling points, with high expression levels in siliques and seeds, whereas no expression of BnKIX8.A1 or BnKIX8.C1 was detected. (2) Homozygous T-DNA-free single mutants, bnkix8.a7 and bnkix8.c7, and the double mutant bnkix8.a7bnkix8.c7 were successfully obtained through CRISPR-Cas9-mediated gene editing. Phenotypic analysis showed that, compared with the wild type, the bnkix8s mutants had significantly increased flower size, silique length, seed size, and 1000-seed weight, as well as significantly higher seed germination and vigor indices. The double mutant showed an additive phenotype, indicating functional redundancy between BnKIX8.A7 and BnKIX8.C7 in regulating rapeseed growth and development. (3) Evaluation of S. sclerotiorum resistance showed that, at 36 and 48 h post-inoculation, lesion areas in the bnkix8s mutants were significantly larger than those in the wild type, suggesting that BnKIX8.A7 and BnKIX8.C7 positively regulate resistance to S. sclerotiorum. (4) KEGG enrichment analysis of transcriptome data indicated that, under normal growth conditions, BnKIX8.A7 and BnKIX8.C7 may contribute to the growth–defense balance by influencing IAA and JA signal transduction pathways. At 24 h post-inoculation with S. sclerotiorum, BnKIX8.A7 and BnKIX8.C7 may participate in the resistance response by affecting α-linolenic acid metabolism and thiamine metabolism pathways. This study provides candidate pathways and genetic resources for further elucidating the functions of BnKIX8s and offers a theoretical basis for breeding high-yield and disease-resistant rapeseed varieties.

Key words: Brassica napus, BnKIX8, CRISPR-Cas9, Sclerotinia sclerotiorum, transcriptome

罗丽娅, 马伍慧, 杨龙, 祝培, 李艳, 许本波, 张学昆, 徐劲松. BnKIX8调控甘蓝型油菜器官大小与菌核病抗性的初步研究[J]. 作物学报, 2026, 52(11): 3334-3351.

Luo Li-Ya, Ma Wu-Hui, Yang Long, Zhu Pei, Li Yan, Xu Ben-Bo, Zhang Xue-Kun, Xu Jin-Song. Preliminary study on the regulation of organ size and Sclerotinia sclerotiorum resistance by BnKIX8 in rapeseed (Brassica napus L.)[J]. Acta Agronomica Sinica, 2026, 52(11): 3334-3351.

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://zwxb.chinacrops.org/CN/10.3724/SP.J.1006.2026.65014

https://zwxb.chinacrops.org/CN/Y2026/V52/I11/3334

参考文献

[1] 刘成, 冯中朝, 肖唐华, 等. 我国油菜产业发展现状、潜力及对策. 中国油料作物学报, 2019, 41: 485–489.
Liu C, Feng Z C, Xiao T H, et al. Development, potential and adaptation of Chinese rapeseed industry. Chin J Oil Crop Sci, 2019, 41: 485–489 (in Chinese with English abstract).

[2] Shi J Q, Li R Y, Qiu D, et al. Unraveling the complex trait of crop yield with quantitative trait loci mapping in Brassica napus. Genetics, 2009, 182: 851–861.

[3] Moles A T, Westoby M. Seedling survival and seed size: a synthesis of the literature. J Ecol, 2004, 92: 372–383.

[4] 曹维, 赵静, 禹艳坤, 等. 调控植物种子大小的分子机制综述. 江苏农业科学, 2020, 48(6): 1–7.
Cao W, Zhao J, Yu Y K, et al. Molecular mechanisms of regulating plant seed size: a review. Jiangsu Agric Sci, 2020, 48(6): 1–7 (in Chinese with English abstract).

[5] Li N, Xu R, Li Y H. Molecular networks of seed size control in plants. Annu Rev Plant Biol, 2019, 70: 435–463.

[6] Thakur J K, Yadav A, Yadav G. Molecular recognition by the KIX domain and its role in gene regulation. Nucleic Acids Res, 2014, 42: 2112–2125.

[7] White D W R. PEAPOD regulates Lamina size and curvature in Arabidopsis. Proc Natl Acad Sci U S A, 2006, 103: 13238–13243.

[8] Gonzalez N, Pauwels L, Baekelandt A, et al. A repressor protein complex regulates leaf growth in Arabidopsis. Plant Cell, 2015, 27: 2273–2287.

[9] Baekelandt A, Pauwels L, Wang Z B, et al. Arabidopsis leaf flatness is regulated by PPD2 and NINJA through repression of CYCLIN D3 genes. Plant Physiol, 2018, 178: 217–232.

[10] Liu Z P, Li N, Zhang Y Y, et al. Transcriptional repression of GIF1 by the KIX-PPD-MYC repressor complex controls seed size in Arabidopsis. Nat Commun, 2020, 11: 1846.

[11] Swinnen G, Mauxion J P, Baekelandt A, et al. SlKIX8 and SlKIX9 are negative regulators of leaf and fruit growth in tomato. Plant Physiol, 2022, 188: 382–396.

[12] Mao Y W, Zhou S L, Yang J, et al. The MIO1-MtKIX8 module regulates the organ size in Medicago truncatula. Physiol Plant, 2023, 175: e14046.

[13] Nguyen C X, Paddock K J, Zhang Z Y, et al. GmKIX8-1 regulates organ size in soybean and is the causative gene for the major seed weight QTL qSw17-1. New Phytol, 2021, 229: 920–934.

[14] Li X, Liu W, Zhuang L L, et al. BIGGER ORGANS and ELEPHANT EAR-LIKE LEAF1 control organ size and floral organ internal asymmetry in pea. J Exp Bot, 2019, 70: 179–191.

[15] Huot B, Yao J, Montgomery B L, et al. Growth-defense tradeoffs in plants: a balancing act to optimize fitness. Mol Plant, 2014, 7: 1267–1287.

[16] He Z H, Webster S, He S Y. Growth-defense trade-offs in plants. Curr Biol, 2022, 32: R634–R639.

[17] Wei L J, Jian H J, Lu K, et al. Genome-wide association analysis and differential expression analysis of resistance to Sclerotinia stem rot in Brassica napus. Plant Biotechnol J, 2016, 14: 1368–1380.

[18] Yan L H, Wei S W, Wu Y R, et al. High-efficiency genome editing in Arabidopsis using YAO promoter-driven CRISPR/Cas9 system. Mol Plant, 2015, 8: 1820–1823.

[19] 中华人民共和国国家市场监督管理总局, 中国国家标准化管理委员会. 农作物种子检验规程第4部分: 播种质量发芽试验: GB/T 3543.4-2025. 北京: 中国标准出版社, 2025.
State Administration of the People’s Republic of China for Market Regulation, Standardization Administration of the People’s Republic of China. Rules for agricultural seed testing: Part 4: Sowing Quality–Germination Test: GB/T 3543.4-2025. Beijing: Standards Press of China, 2025 (in Chinese).

[20] 何烈干, 宋来强, 汤洁, 等. 油菜菌核病抗性鉴定方法比较及抗病种质资源的筛选. 江苏农业科学, 2018, 46(18): 90–93.
He L G, Song L Q, Tang J, et al. Comparison of identification methods for resistance to Sclerotinia sclerotiorum and screening of resistant materials of rapeseed. Jiangsu Agric Sci, 2018, 46(18): 90–93 (in Chinese with English abstract).

[21] 黎家, 李传友. 新中国成立70年来植物激素研究进展. 中国科学(生命科学), 2019, 49: 1227–1281.
Li J, Li C Y. Seventy-year major research progress in plant hormones by Chinese scholars. Sci China Ser C, 2019, 49: 1227–1281 (in Chinese with English abstract).

[22] Waadt R, Seller C A, Hsu P K, et al. Plant hormone regulation of abiotic stress responses. Nat Rev Mol Cell Biol, 2022, 23: 680–694.

[23] Major I T, Yoshida Y, Campos M L, et al. Regulation of growth–defense balance by the JASMONATE ZIM-DOMAIN (JAZ)-MYC transcriptional module. New Phytol, 2017, 215: 1533–1547.

[24] Fernández-Calvo P, Chini A, Fernández-Barbero G, et al. The Arabidopsis bHLH transcription factors MYC3 and MYC4 are targets of JAZ repressors and act additively with MYC2 in the activation of jasmonate responses. Plant Cell, 2011, 23: 701–715.

[25] Thines B, Katsir L, Melotto M, et al. JAZ repressor proteins are targets of the SCFCOI1 complex during jasmonate signalling. Nature, 2007, 448: 661–665.

[26] Wasternack C, Song S S. Jasmonates: biosynthesis, metabolism, and signaling by proteins activating and repressing transcription. J Exp Bot, 2017, 68: 1303–1321.

[27] Goyer A. Thiamine in plants: aspects of its metabolism and functions. Phytochemistry, 2010, 71: 1615–1624.

[28] Julliard J H, Douce R. Biosynthesis of the thiazole moiety of thiamin (vitamin B1) in higher plant chloroplasts. Proc Natl Acad Sci USA, 1991, 88: 2042–2045.

[29] Ahn I P, Kim S, Lee Y H, et al. Vitamin B1-induced priming is dependent on hydrogen peroxide and the NPR1 gene in Arabidopsis. Plant Physiol, 2007, 143: 838–848.

[30] 何晓玲, 刘鹏程, 马伯军, 等. 基于CRISPR/Cas9的基因编辑技术研究进展及其在植物中的应用. 植物学报, 2022, 57: 508–531.
He X L, Liu P C, Ma B J, et al. Advance in gene-editing technology based on CRISPR/Cas9 and its application in plants. Bull Bot, 2022, 57: 508–531 (in Chinese with English abstract).

[31] 许磊, 何菡子, 范楚川. 基于CRISPR/Cas9的基因编辑及其在甘蓝型油菜中的应用. 华中农业大学学报, 2024, 43(5): 51–64.
Xu L, He H Z, Fan C C. CRISPR/Cas9 based gene editing and its application in Brassica napus L. J Huazhong Agric Univ, 2024, 43(5): 51–64 (in Chinese with English abstract).

[32] Du M M, Spalding E P, Gray W M. Rapid auxin-mediated cell expansion. Annu Rev Plant Biol, 2020, 71: 379–402.

[33] Browse J. Jasmonate passes muster: a receptor and targets for the defense hormone. Annu Rev Plant Biol, 2009, 60: 183–205.

[34] Sood M. Jasmonates: “the master switch” for regulation of developmental and stress responses in plants. J Plant Growth Regul, 2023, 42: 5247–5265.

[35] Roychowdhury R, Hada A, Biswas S, et al. Jasmonic acid (JA) in plant immune response: unravelling complex molecular mechanisms and networking of defence signalling against pathogens. J Plant Growth Regul, 2025, 44: 89–114.

[36] Sun S H, Han X Y, Zhang Y Y, et al. Exogenous thiamine regulates multiple plant immunity-associated pathways to inhibit MCMV infection. J Agric Food Chem, 2025, 73: 14353–14362.

[37] Feng M, Liu Y, Zhao Y, et al. Genome-wide analysis of the thiamine biosynthesis gene families in common bean reveals their crucial roles conferring resistance to Fusarium Wilt. Biology, 2025, 14: 1366

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

BnKIX8调控甘蓝型油菜器官大小与菌核病抗性的初步研究

Preliminary study on the regulation of organ size and Sclerotinia sclerotiorum resistance by BnKIX8 in rapeseed (Brassica napus L.)

PDF

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 10

关于本刊

在线期刊

作者中心

相关单位

联系我们

[1]	金海润, 欧阳青静, 龙旭, 晏中彬, 鲁海瑞, 古开烨, 田恩堂. 甘蓝型油菜种质资源菌核病抗性鉴定与候选基因筛选[J]. 作物学报, 2026, 52(7): 2027-2044.
[2]	黄焰新, 闫旭宇, 曹艳, 卫怡涵, 张佳宁, 汪奕彤, 李玲. 外源钙协同褪黑素缓解棉花幼苗镉胁迫的生理效应及转录代谢调控分析[J]. 作物学报, 2026, 52(7): 2159-2179.
[3]	王亚, 赵宜婷, 王宙, 杨俊芳, 张宏斌, 曹越. 转录组-代谢组联合分析蓖麻蜡质合成相关基因[J]. 作物学报, 2026, 52(6): 1774-1787.
[4]	陈雪燕, 何华川, 李政嘉, 董新盼, 李藕琪, 刘小云, 李丹萍, 陈志伟, 刘国霞, 吕胜源, 吴印莹, 赵振东, 曹新有, 万何平. 水培盐碱复合胁迫下‘济麦60’苗期根系有机酸分泌动态变化及其转录调控机制[J]. 作物学报, 2026, 52(6): 1859-1875.
[5]	邹仪妹, 徐敏, 汪海洋, 姚辉, 王加峰, 刘浩, 任代胜. 两系不育系水稻幼苗根系响应盐胁迫的转录因子调控网络鉴定[J]. 作物学报, 2026, 52(6): 1728-1742.
[6]	蔡兆琴, 何观咏, 何文, 阮丽霞, 梁振华, 李永珍, 李恒锐, 陈会鲜. 木薯分枝发育过程的动态转录组分析与关键基因发掘[J]. 作物学报, 2026, 52(5): 1430-1441.
[7]	张曦, 王广恩, 李邵琦, 刘祎, 李俊兰, 钱玉源. 基于转录组测序解析陆海杂交姊妹系马克隆值差异的形成机制[J]. 作物学报, 2026, 52(5): 1442-1458.
[8]	韩亚鑫, 何冠华, 张小琼, 张登峰, 李永祥, 刘旭洋, 王天宇, 黎裕, 邹华文, 李春辉. 基于RNA-Seq和BSA-Seq联合分析挖掘玉米侧根密度基因资源[J]. 作物学报, 2026, 52(5): 1341-1352.
[9]	杨亚莉, 徐明睿, 马越飞, 海艺蕊, 刘凯栋, 刘万茂, 孙颖. 玉米根尖及整根响应缺铁的转录组比较研究[J]. 作物学报, 2026, 52(4): 1006-1021.
[10]	马亮, 马璐, 张舒钰, 章慧敏, 王仁明, 宋旭东, 张振良, 冒宇翔, 陆虎华, 陈国清, 郝德荣, 周广飞. 玉米苞叶数目转录组分析及候选基因鉴定[J]. 作物学报, 2026, 52(3): 790-801.
[11]	杨宗桃, 杨婷, 王禹童, 艾静, 李燕烨, 刘家勇, 邓军, 赵勇, 张跃彬. 甘蔗CLC基因家族鉴定与表达分析[J]. 作物学报, 2026, 52(3): 722-734.
[12]	张宇, 刘芳, 蔡诚诚, 杨小华, 吉阿么石扎, 杨元军, 王西瑶. 溴乙烷与赤霉素协同处理破除马铃薯块茎休眠的机理初探[J]. 作物学报, 2026, 52(3): 825-838.
[13]	于永超, 刘明, 靳容, 赵鹏, 张强强, 王静, 朱晓亚, 唐忠厚. 甘薯高氮徒长的生理机制和转录组分析研究[J]. 作物学报, 2026, 52(3): 813-824.
[14]	张力岚, 杨军, 王让剑. 基于WGCNA发掘茶树糖苷类香气前体含量性状相关的候选基因[J]. 作物学报, 2026, 52(2): 494-513.
[15]	马毅娜, 吴晓明玉, 李藕琪, 王圆, 陈丽, 张盈川, 赵伦, 文静, 傅廷栋, 沈金雄. Bna-miR1040-EIF3A模块调控油菜开花时间的功能研究[J]. 作物学报, 2026, 52(2): 349-362.