欢迎访问作物学报，今天是
作物学报

作物学报 ›› 2024, Vol. 50 ›› Issue (8): 1896-1906.doi: 10.3724/SP.J.1006.2024.43007

所属专题： 玉米：遗传育种·种质资源·分子遗传学

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

基于大刍草渗入系的玉米抗旱优异等位基因挖掘

刘爽(), 李珅, 王东梅, 沙小茜, 何冠华, 张登峰, 李永祥, 刘旭洋, 王天宇, 黎裕, 李春辉()   

  1. 作物基因资源与育种全国重点实验室 / 中国农业科学院作物科学研究所, 北京 100081
  • 收稿日期:2024-01-24 接受日期:2024-04-01 出版日期:2024-08-12 网络出版日期:2024-04-22
  • 通讯作者: * 李春辉, E-mail: lichunhui@caas.cn
  • 作者简介:E-mail: 1971556590@qq.com
  • 基金资助:
    财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-02-03);中国农业科学院科技创新工程项目(CAAS-ZDRW202004)

Superior allele genes mining for drought tolerance in maize based on introgression line from a cross between maize and teosinte

LIU Shuang(), LI Shen, WANG Dong-Mei, SHA Xiao-Qian, HE Guan-Hua, ZHANG Deng-Feng, LI Yong-Xiang, LIU Xu-Yang, WANG Tian-Yu, LI Yu, LI Chun-Hui()   

  1. State Key Laboratory of Crop Gene Resources and Breeding / Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2024-01-24 Accepted:2024-04-01 Published:2024-08-12 Published online:2024-04-22
  • Contact: * E-mail: lichunhui@caas.cn
  • Supported by:
    China Agriculture Research System of MOF and MARA(CARS-02-03);Innovation Program of Chinese Academy of Agricultural Sciences(CAAS-ZDRW202004)

RichHTML

38

PDF

626

摘要：

干旱是影响玉米生产的主要非生物胁迫之一。为了挖掘玉米优异抗旱基因, 本研究基于墨西哥大刍草和玉米自交系PH4CV构建的BC2F6群体, 通过苗期抗旱性初步鉴定, 筛选出抗旱性强的渗入系TP180。干旱胁迫后发现渗入系TP180较轮回亲本PH4CV萎蔫程度更小, 并且复水后存活率显著高于PH4CV。全基因组基因型鉴定发现, 渗入系TP180含有0.6%的墨西哥大刍草导入。通过开展TP180和PH4CV不同水分条件下的转录组测序, 在渗入系TP180和PH4CV之间共鉴定了 2307个差异表达基因, 不同胁迫重叠差异表达基因共122个, 这些基因涉及生长素途径, 茉莉酸途径等, 包含多个转录因子。整合差异表达基因和含大刍草导入区段分析, 鉴定出2个抗旱候选基因Zm00001d033050和Zm00001d002025, 并进一步开展了RT-PCR分析。本研究为挖掘大刍草中抗旱基因资源提供了重要材料和信息基础。

关键词: 玉米, 大刍草, 渗入系, 抗旱, 转录组分析

Abstract:

Drought is one of the major abiotic stresses affecting maize production. In order to explore new genes for drought tolerance in maize, the BC2F6 population constructed on the basis of teosinte and PH4CV was screened for the drought-tolerant introgression lines TP180 through the preliminary identification of drought tolerance at seedling stage. After drought stress, the TP180 was less wilting than its recurrent parent PH4CV, and the survival rate of TP180 was significantly higher than PH4CV after rehydration. Genome-wide genotypic identification revealed that introgression line TP180 contained 0.6% of the teosinte genome. By transcriptomic analysis of TP180 and PH4CV under different water conditions, a total of 2307 differentially expressed genes were identified between TP180 and PH4CV, and 122 of the differentially expressed genes were identified under both two drought stresses conditions. These genes were related to the growth hormone pathway, jasmonic acid pathway, etc., and contained multiple transcription factors. Integrating the differentially expressed genes and the analysis of introgression region containing the teosinte genome, two drought-resistant candidate genes (Zm00001d033050 and Zm00001d002025) were identified and further validated by RT-PCR. This study provides an important germplasm and information for mining drought-resistant gene of the teosinte.

Key words: maize, teosinte, introgression line, drought tolerance, transcriptomic analysis

表1

本研究所使用的引物信息表"

引物名称Primer name 引物序列Primer sequence (5'-3')
GAPDH-F CCCTTCATCACCACGGACTAC
GAPDH-R AACCTTCTTGGCACCACCCT
ReTIFY9-F CAGCGGAACGTGCAGAAACA
ReTIFY9-R2 GCCTTGCGCTTCTCCATGAAC
ReEreb24-F CACCACTGCTACGCCAATG
ReEreb24-R ATTGAAAGCGACCGGGGTG

图1

TP180和PH4CV抗旱性分析与干旱处理存活率分析 A: 正常水分; B: 中度干旱胁迫; C: 重度干旱胁迫; D: 复水; E: 存活率。**表示在P < 0.01水平差异显著。"

图2

大刍草导入区段分析"

图3

不同处理下差异表达基因数目 A: Venn图描述所有差异表达基因的数量; B: 上调、下调的差异基因总数。WW: 正常水分; MD: 中度干旱胁迫; SD: 重度干旱胁迫。"

图4

不同处理下差异表达基因GO富集 A: 中度干旱胁迫上调表达基因; B: 中度干旱胁迫下调表达基因; C: 重度干旱胁迫上调表达基因; D: 重度干旱胁迫中度干旱胁迫下调表达基因。"

图5

差异表达中已知抗旱基因不同处理下表达量 FPKM代表差异显著基因的表达水平, WW代表正常水分(watered)条件下基因表达量, MD代表中度干旱胁迫(moderate-drought)条件下基因表达量, SD代表重度干旱胁迫(severe-drought)条件下基因表达量。*和**分别表示在P < 0.05和P < 0.01水平差异显著, ns表示无差异显著性。"

图6

候选抗旱基因不同处理下基因表达水平 A~B图为转录组数据, C~D图为RT-PCR实验结果, FPKM代表差异显著基因的表达量, Relative expression level为RT-PCR实验得到的基因表达量, WW代表正常水分(watered)条件下基因表达量, MD代表中度干旱胁迫(moderate-drought)条件下基因表达量, SD代表重度干旱胁迫(severe-drought)条件下基因表达量。*和**分别表示在P < 0.05和P < 0.01水平差异显著, ns表示无差异显著性。"

[1] McMillen M S, Mahama A A, Sibiya J, Lübberstedt T, Suza W P. Improving drought tolerance in maize: tools and techniques. Front Genet, 2022, 13: 1001001.
[2] Li P, Zhang Y, Yin S, Zhu P, Pan T, Xu Y, Wang J, Hao D, Fang H, Xu C, Yang Z. QTL-by-environment interaction in the response of maize root and shoot traits to different water regimes. Front Plant Sci, 2018, 9: 229.
doi: 10.3389/fpls.2018.00229 pmid: 29527220
[3] Li C, Sun B, Li Y, Liu C, Wu X, Zhang D, Shi Y, Song Y, Buckler ES, Zhang Z, Wang T, Li Y. Numerous genetic loci identified for drought tolerance in the maize nested association mapping populations. BMC Genomics, 2016, 17: 894.
pmid: 27825295
[4] Riedelsheimer C, Lisec J, Czedik-Eysenberg A, Sulpice R, Flis A, Grieder C, Altmann T, Stitt M, Willmitzer L, Melchinger A E. Genome-wide association mapping of leaf metabolic profiles for dissecting complex traits in maize. Proc Natl Acad Sci USA, 2012, 109: 8872-8877.
doi: 10.1073/pnas.1120813109 pmid: 22615396
[5] Liu S, Qin F. Genetic dissection of maize drought tolerance for trait improvement. Mol Breed, 2021, 41: 8.
[6] Wang X, Wang H, Liu S, Ferjani A, Li J, Yan J, Yang X, Qin F. Genetic variation in ZmVPP1 contributes to drought tolerance in maize seedlings. Nat Genet, 2016, 48: 1233-1241.
[7] Mao H, Wang H, Liu S, Li Z, Yang X, Yan J, Li J, Tran L S, Qin F. A transposable element in a NAC gene is associated with drought tolerance in maize seedlings. Nat Commun, 2015, 6: 8326.
[8] Li C, Guo J, Wang D, Chen X, Guan H, Li Y, Zhang D, Liu X, He G, Wang T, Li Y. Genomic insight into changes of root architecture under drought stress in maize. Plant Cell Environ, 2023, 46: 1860-1872.
[9] Sun X, Xiang Y, Dou N, Zhang H, Pei S, Franco A V, Menon M, Monier B, Ferebee T, Liu T, Liu S, Gao Y, Wang J, Terzaghi W, Yan J, Hearne S, Li L, Li F, Dai M. The role of transposon inverted repeats in balancing drought tolerance and yield-related traits in maize. Nat Biotechnol, 2023, 41: 120-127.
[10] Yang N, Wang Y, Liu X, Jin M, Vallebueno-Estrada M, Calfee E, Chen L, Dilkes B P, Gui S, Fan X, Harper T K, Kennett D J, Li W, Lu Y, Ding J, Chen Z, Luo J, Mambakkam S, Menon M, Snodgrass S, Veller C, Wu S, Wu S, Zhuo L, Xiao Y, Yang X, Stitzer M C, Runcie D, Yan J, Ross-Ibarra J. Two teosintes made modern maize. Science, 2023, 382: eadg8940.
[11] Chen L, Luo J, Jin M, Yang N, Liu X, Peng Y, Li W, Phillips A, Cameron B, Bernal J S, Rellán-Álvarez R, Sawers R J H, Liu Q, Yin Y, Ye X, Yan J, Zhang Q, Zhang X, Wu S, Gui S, Wei W, Wang Y, Luo Y, Jiang C, Deng M, Jin M, Jian L, Yu Y, Zhang M, Yang X, Hufford M B, Fernie A R, Warburton M L, Ross-Ibarra J, Yan J. Genome sequencing reveals evidence of adaptive variation in the genus Zea. Nat Genet, 2022, 54: 1736-1745.
doi: 10.1038/s41588-022-01184-y pmid: 36266506
[12] 刘杰, 严建兵. 大刍草稀有等位基因促进玉米密植高产. 植物学报, 2019, 54: 554-557.
doi: 10.11983/CBB19119
Liu J, Yan J B. A teosinte rare allele increases maize plant density and yield. Chin Bull Bot, 2019, 54: 554-557 (in Chinese with English abstract).
[13] Feng X, Jia L, Cai Y, Guan H, Zheng D, Zhang W, Xiong H, Zhou H, Wen Y, Hu Y, Zhang X, Wang Q, Wu F, Xu J, Lu Y. ABA- inducible DEEPER ROOTING 1improves adaptation of maize to water deficiency. Plant Biotechnol J, 2022, 20: 2077-2088.
[14] Wang K, Zhang Z, Sha X, Yu P, Li Y, Zhang D, Liu X, He G, Li Y, Wang T, Guo J, Chen J, Li C. Identification of a new QTL underlying seminal root number in a maize-teosinte population. Front Plant Sci, 2023, 14: 1132017.
[15] Chen G, Liu C, Gao Z, Zhang Y, Jiang H, Zhu L, Ren D, Yu L, Xu G, Qian Q. OsHAK1, a high-affinity potassium transporter, positively regulates responses to drought stress in rice. Front Plant Sci, 2017, 8: 1885.
doi: 10.3389/fpls.2017.01885 pmid: 29163608
[16] Naithani S, Dikeman D, Garg P, Al-Bader N, Jaiswal P. Beyond gene ontology (GO): using biocuration approach to improve the gene nomenclature and functional annotation of rice S-domain kinase subfamily. PeerJ, 2021, 9: e11052.
[17] Zhang P Y, Qiu X, Fu J X, Wang G R, Wei L, Wang T C. Systematic analysis of differentially expressed ZmMYB genes related to drought stress in maize. Physiol Mol Biol Plants, 2021, 27: 1295-1309.
[18] Qu X, Zou J, Wang J, Yang K, Wang X, Le J. A rice R2R3-type MYB transcription factor OsFLP positively regulates drought stress response via OsNAC. Int J Mol Sci, 2022, 23: 5873.
[19] Zhao P X, Miao Z Q, Zhang J, Chen S Y, Liu Q Q, Xiang C B. Arabidopsis MADS-box factor AGL16 negatively regulates drought resistance via stomatal density and stomatal movement. J Exp Bot, 2020, 71: 6092-6106.
[20] Song W, Zhao H, Zhang X, Lei L, Lai J. Genome-wide identification of VQ motif-containing proteins and their expression profiles under abiotic stresses in maize. Front Plant Sci, 2016, 6: 1177.
[21] Isokpehi R D, Simmons S S, Cohly H H, Ekunwe S I, Begonia G B, Ayensu W K. Identification of drought-responsive universal stress proteins in viridiplantae. Bioinform Biol Insights, 2011, 5: 41-58.
[22] Zhang F, Wu J, Sade N, Wu S, Egbaria A, Fernie A R, Yan J, Qin F, Chen W, Brotman Y, Dai M. Genomic basis underlying the metabolome-mediated drought adaptation of maize. Genome Biol, 2021, 22: 260.
doi: 10.1186/s13059-021-02481-1 pmid: 34488839
[23] Gao H, Cui J, Liu S, Wang S, Lian Y, Bai Y, Zhu T, Wu H, Wang Y, Yang S, Li X, Zhuang J, Chen L, Gong Z, Qin F. Natural variations of ZmSRO1d modulate the trade-off between drought resistance and yield by affecting ZmRBOHC-mediated stomatal ROS production in maize. Mol Plant, 2022, 15: 1558-1574.
[24] Yang Y, Wang B, Wang J, He C, Zhang D, Li P, Zhang J, Li Z. Transcription factors ZmNF-YA1 and ZmNF-YB16 regulate plant growth and drought tolerance in maize. Plant Phys, 2022, 190: 1506-1525.
[25] Zhao W, Huang H, Wang J, Wang X, Xu B, Yao X, Sun L, Yang R, Wang J, Sun A, Wang S. Jasmonic acid enhances osmotic stress responses by MYC2-mediated inhibition of protein phosphatase 2C1 and response regulators 26 transcription factor in tomato. Plant J, 2023, 113: 546-561.
[26] Nie J, Wen C, Xi L, Lv S, Zhao Q, Kou Y, Ma N, Zhao L, Zhou X. The AP2/ERF transcription factor CmERF053 of chrysanthemum positively regulates shoot branching, lateral root, and drought tolerance. Plant Cell Rep, 2018, 37: 1049-1060.
doi: 10.1007/s00299-018-2290-9 pmid: 29687169
[27] 陈小晶, 王东梅, 关红辉, 郭剑, 沙小茜, 李永祥, 张登峰, 刘旭洋, 何冠华, 石云素, 宋燕春, 王天宇, 黎裕, 刘颖慧, 李春辉. 玉米CIPK基因家族的鉴定及ZmCIPK3的抗旱性功能研究. 植物遗传资源学报, 2022, 23: 1064-1075.
doi: 10.13430/j.cnki.jpgr.20220107006
Chen X J, Wang D M, Guan H H, Guo J, Sha X Q, Li Y X, Zhang D F, Liu X Y, He G H, Shi Y S, Song Y C, Wang T Y, Li Y, Liu Y H, Li C H. Identification of CIPK gene family members and investigation of the drought tolerance of ZmCIPK3 in maize. J Plant Genet Resour, 2022, 23: 1064-1075 (in Chinese with English abstract).
[28] Tian J, Wang C, Xia J, Wu L, Xu G, Wu W, Li D, Qin W, Han X, Chen Q, Jin W, Tian F. Teosinte ligule allele narrows plant architecture and enhances high-density maize yields. Science, 2019, 365: 658-664.
doi: 10.1126/science.aax5482 pmid: 31416957
[29] Depuydt S, Hardtke C S. Hormone signalling crosstalk in plant growth regulation. Curr Biol, 2011, 21: R365-373.
[30] 艾蓉, 张春, 悦曼芳, 邹华文, 吴忠义. 玉米转录因子ZmEREB211对非生物逆境胁迫的应答. 作物学报, 2023, 49: 2433-2445.
doi: 10.3724/SP.J.1006.2023.23071
Ai R, Zhang C, Yue M F, Zou H W, Wu Z Y. Response of maize transcriptional factor ZmEREB211 to abiotic stress. Acta Agron Sin, 2023, 49: 2433-2445 (in Chinese with English abstract).
[31] 王丽平, 王晓钰, 傅竞也, 王强. 玉米转录因子ZmMYB12提高植物抗旱性和低磷耐受性的功能鉴定. 作物学报, 2024, 50: 76-88.
doi: 10.3724/SP.J.1006.2024.33007
Wang L P, Wang X Y, Fu J Y, Wang Q. Functional identification of maize transcription factor ZmMYB12 to enhance drought resistance and low phosphorus tolerance in plants. Acta Agron Sin, 2024, 50: 76-88 (in Chinese with English abstract).
[32] Xiang Y, Sun X, Bian X, Wei T, Han T, Yan J, Zhang A. The transcription factor ZmNAC49 reduces stomatal density and improves drought tolerance in maize. J Exp Bot, 2021, 72: 1399-1410.
doi: 10.1093/jxb/eraa507 pmid: 33130877
[33] Zhang Z, Li X, Yu R, Han M, Wu Z. Isolation, structural analysis, and expression characteristics of the maize TIFY gene family. Mol Genet Genomics, 2015, 290: 1849-1858.
doi: 10.1007/s00438-015-1042-6 pmid: 25862669
[34] Ye H, Du H, Tang N, Li X, Xiong L. Identification and expression profiling analysis of TIFY family genes involved in stress and phytohormone responses in rice. Plant Mol Biol, 2009, 71: 291-305.
doi: 10.1007/s11103-009-9524-8 pmid: 19618278
[35] Qi H, Liang K, Ke Y, Wang J, Yang P, Yu F, Qiu F. Advances of apetala2/ethylene response factors in regulating development and stress response in maize. Int J Mol Sci, 2023, 24: 5416.
[36] Zhu Y, Liu Y, Zhou K, Tian C, Aslam M, Zhang B, Liu W, Zou H. Overexpression of ZmEREBP60 enhances drought tolerance in maize. J Plant Physiol, 2022, 275: 153763.
[1] 刘恩波, 陈静, 李红星, 于宁宁, 任佰朝, 赵斌, 刘鹏, 张吉旺. 遮阴改变源-库平衡和调节碳水化合物代谢进而抑制夏玉米幼穗发育[J]. 作物学报, 2026, 52(6): 1891-1901.
[2] 梁进宇, 尹嘉德, 王红丽, 张国平, 侯慧芝, 董博, 马明生. 基于无人机高光谱和机器学习的旱地饲用玉米叶片氮含量估测[J]. 作物学报, 2026, 52(6): 1788-1801.
[3] 孙淑凤, 许振南, 黄嘉鑫, 翁建峰, 李新海. 玉米MAPK家族全基因组鉴定及其对拟轮枝镰孢菌感染的响应[J]. 作物学报, 2026, 52(5): 1291-1308.
[4] 张宁宁, 滕雨菲, 任娜娜, 魏兴卓, 闫书豪, 樊可心, 王永宏, 陈文康, 张兴华, 朱万超, 徐淑兔, 薛吉全. 201份玉米自交系抗旱表型评价及可塑性分析[J]. 作物学报, 2026, 52(5): 1309-1325.
[5] 杨欣雨, 崔文涛, 迪力尼格尔·阿力木, 汪凯翔, 吴鹏昊, 任姣姣. 玉米穗上叶片数全基因组关联分析和全基因组选择[J]. 作物学报, 2026, 52(5): 1573-1590.
[6] 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.
[7] 杨扬, 常诗惠, 田红丽, 易红梅, 王璐, 任洁, 范亚明, 刘亚维, 王凤格, 赵久然. 不同生态区国审玉米品种的遗传多样性分析[J]. 作物学报, 2026, 52(5): 1352-1364.
[8] 张鸿蓉, 王菲儿, 李盼, 仇海龙, 朱静, 赵连豪, 南运有, 何蔚, 樊志龙, 胡发龙, 柴强, 殷文. 减量20%灌水与25%有机肥替代化肥提高青贮玉米产量的光合特性[J]. 作物学报, 2026, 52(5): 1487-1500.
[9] 蔡宏玮, 于爱忠, 姜科强, 王鹏飞, 王玉珑, 霍建喆, 庞小能, 尹波, 尚永盼. 干旱灌区有机肥替代部分化肥促进甜玉米产量提升的关键机制[J]. 作物学报, 2026, 52(4): 1166-1180.
[10] 田红丽, 杨扬, 范亚明, 易红梅, 郭丹丹, 王凤格, 赵久然. 适于玉米品种鉴定的一套三等位变异SNP新型标记组合[J]. 作物学报, 2026, 52(4): 993-1005.
[11] 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.
[12] 张超, 郭欢, 李忠玲, 岳淑宁, 赵娜. 基于BSA-seq技术定位玉米籽粒花青素关联基因[J]. 作物学报, 2026, 52(3): 780-789.
[13] 郭向阳, 涂亮, 王栋, 刘鹏飞, 王安贵, 易强, 任洪, 李刚, 祝云芳, 吴迅, 蒋喻林, 田丰, 陈泽辉. 热带Suwan种质在我国玉米种质改良中的创新与利用[J]. 作物学报, 2026, 52(3): 655-664.
[14] 孟成, 王哲. 玉米ZmPFK基因家族全基因组鉴定及响应胁迫表达分析[J]. 作物学报, 2026, 52(3): 764-779.
[15] 李新浩, 邢梦柯, 周梓惠, 李思烨, 任昊, 王洪章, 赖华江. 外源褪黑素通过协调光反应与暗反应增强玉米苗期的耐热性[J]. 作物学报, 2026, 52(3): 839-856.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备15008789号-2