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作物学报 ›› 2025, Vol. 51 ›› Issue (10): 2570-2580.doi: 10.3724/SP.J.1006.2025.54038

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

棉花减数分裂进程鉴定体系构建及其对高温胁迫的响应分析

李亚玮(), 徐盈盈, 左春阳, 刘若男, 梁亚军, 孔杰, 张献龙, 闵玲()   

  1. 华中农业大学作物遗传改良全国重点实验室 / 湖北洪山实验室, 湖北武汉 430070
  • 收稿日期:2025-03-20 接受日期:2025-07-09 出版日期:2025-10-12 网络出版日期:2025-07-15
  • 通讯作者: *闵玲, E-mail: lingmin@mail.hzau.edu.cn
  • 作者简介:李亚玮, E-mail: liyawei@webmail.hzau.edu.cn
    **同等贡献
  • 基金资助:
    新疆维吾尔自治区2023年“揭榜挂帅”棉花品种重大农艺性状解析与分子设计育种项目(2023A01-3);新疆人才发展基金“优质耐高温棉花关键基因发掘与育种应用”项目(202403-03);湖北省杰出青年基金-棉花响应高温胁迫遗传机制解析及育种应用项目(2024AFA059)

Construction of a meiotic progression identification system in cotton and analysis of its response to high-temperature stress

LI Ya-Wei(), XU Ying-Ying, ZUO Chun-Yang, LIU Ruo-Nan, LIANG Ya-Jun, KONG Jie, ZHANG Xian-Long, MIN Ling()   

  1. National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University / Hubei Hongshan Laboratory, Wuhan 430070, Hubei, China
  • Received:2025-03-20 Accepted:2025-07-09 Published:2025-10-12 Published online:2025-07-15
  • Contact: *E-mail: lingmin@mail.hzau.edu.cn
  • About author:First author contact:**Contributed equally to this work
  • Supported by:
    “Unveiling and Commanding” Major Agronomic Traits and Molecular Design Breeding in Cotton Varieties of Xinjiang Uyghur Autonomous Region(2023A01-3);Projects Sponsored by the Development Fund for Xinjiang Talents “Exploration and Breeding Application of Key Genes for High-Quality, Heat-Resistant Cotton”(202403-03);Hubei Provincial Outstanding Youth Fund-Genetic Mechanism Analysis and Breeding Application of Cotton in Response to High Temperature Stress(2024AFA059)

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摘要：

高温胁迫加剧是导致棉花减数分裂异常及产量下降的核心问题, 本研究以棉花遗传转化受体材料‘Jin 668'为研究对象, 通过对花蕾盐酸酸解、酶解和乙酸酸解体系的摸索, 系统构建棉花减数分裂进程鉴定技术体系, 并对其响应高温胁迫进行分析。通过观察不同长度花蕾的小孢子母细胞和染色体形态, 确定棉花减数分裂进程与花蕾长度的关系, 并详细描述了减数分裂前期I、中期I、后期I、末期I、中期II、后期II和末期II各个时期的细胞和染色体特征。研究发现, 高温胁迫会对棉花减数分裂前期I的各个阶段产生显著影响, 导致染色体凝集异常、配对失败、交叉互换受阻等, 最终导致小孢子形成受阻或发育异常。本研究建立的棉花减数分裂鉴定技术体系, 有效降低了制片过程中杂质的干扰, 为深入研究棉花减数分裂进程及高温胁迫响应机制提供了重要的技术支持, 为棉花耐高温育种和遗传改良提供了新的思路和方法。

关键词: 棉花, 高温胁迫, 减数分裂, 染色体行为, 制片技术

Abstract:

High-temperature stress has become a critical factor disrupting cotton meiosis and reducing yield. In this study, using the cotton transformation receptor material ‘Jin 668', we systematically established a reliable method for identifying meiotic progression in cotton by optimizing protocols for hydrochloric acid hydrolysis, enzymatic digestion, and acetic acid maceration of cotton flower buds. This optimized system effectively minimized interference from impurities during slide preparation. By examining the morphology of microspore mother cells and chromosomes in buds of varying lengths, we clarified the relationship between bud length and meiotic stage, and characterized cellular and chromosomal features across key stages, including prophase I, metaphase I, anaphase I, telophase I, metaphase II, anaphase II, and telophase II. Our results indicate that high-temperature stress significantly affects the early stages of prophase I, leading to abnormal chromosome condensation, failure of homologous pairing, disrupted crossover recombination, and ultimately defective microspore development. The identification system we developed provides a robust technical foundation for further investigation into cotton meiosis and its response to high-temperature stress, offering valuable strategies for breeding heat-tolerant cotton varieties and facilitating genetic improvement efforts.

Key words: cotton, high-temperature stress, meiosis, chromosome behavior, slide preparation technique

图1

陆地棉‘Jin 668'花药发育时期和花蕾长度对应关系图示 A~J分别表示不同长度花蕾对应的半薄切片图片。半薄切片中图注如下, E: 表皮层; Ar: 孢原细胞; 1°P: 初生周缘细胞; 2°P: 次生周缘细胞; 1°Sp: 初生造孢细胞; Sp: 造孢细胞; En: 药室内壁; ML: 中间层; T: 绒毡层; MMC; 花粉母细胞; Msp: 小孢子。纵向标尺=0.5 cm; 横向标尺=50 μm。"

图2

棉花花药减数分裂时期的最佳盐酸酸解时间摸索 A~D为减数分裂间期(A), 减数分裂前期I (B), 减数分裂末期I (C), 四分体时期(D)的花药被1 mol L-1盐酸酸解5 min后的小孢子母细胞镜检图。E~K为四分体时期样品被1 mol L-1盐酸处理的酸解时间摸索体系。标尺=50 μm。"

图3

棉花花蕾长度和性母细胞形态对应关系分析 A: 有丝分裂; B: 减数分裂间期; C: 减数分裂前期I-细线期; D: 减数分裂前期I-偶线期; E: 减数分裂前期I-粗线期; F: 减数分裂前期I-终变期; G: 减数分裂中期I; H: 减数分裂后期I; I: 减数分裂末期I; J: 减数分裂中期II; K: 减数分裂后期II; L: 四分体时期。标尺= 50 μm。"

图4

棉花最适酶解时间摸索 A~E为酶解不同时间后酶解液中的样品情况, 标尺=1.5 cm; F~J为酶解不同时间后样品经100 μL 0.01 mol L-1柠檬酸钠缓冲液(pH 4.5)清洗后的状态, 标尺=1.5 cm; K~O为显微镜下观察到经酶解和0.01 mol L-1柠檬酸钠缓冲液冲洗后的单个花药状态, 标尺=1000 μm; P~T为酶解不同时间花药里的染色体状态, 标尺=20 μm。"

图5

棉花花药减数分裂时期染色体观察最适乙酸酸解体系摸索 A~F展示了棉花花药在不同乙酸浓度下处理3 min后染色体被杂质覆盖的情况。G~J展示了棉花花药在70%乙酸处理不同时间后染色体被杂质覆盖的情况。70%乙酸分别处理3 min (G)、5 min (H)、7 min (I)、10 min (J)、12 min (K)和15 min (L)。标尺=20 μm。"

图6

棉花花药减数分裂全时期染色体进程观察 A: 减数分裂间期; B: 减数分裂前期I-细线期; C: 减数分裂前期I-偶线期; D: 减数分裂前期I-粗线期; E: 减数分裂前期I-双线期; F: 减数分裂前期I-终变期; G: 减数分裂中期I; H: 减数分裂后期I; I: 减数分裂末期I; J: 减数分裂中期II; K: 减数分裂后期II; L: 四分体时期。标尺为右下角图像标尺, 右上角为放大图。标尺=20 μm。"

图7

常温与高温下棉花减数分裂小孢子母细胞观察 A为常温条件下, 减数分裂I时期小孢子母细胞的形态特征; B~D为高温条件下, 减数分裂I时期小孢子母细胞的形态特征; E为常温条件下, 四分体时期小孢子母细胞的形态特征; F~H为高温条件下, 四分体时期小孢子母细胞的形态特征。NT: 常温; HT: 高温。标尺=50 μm。"

图8

常温与高温下棉花减数分裂各阶段染色体形态变化观察 A和G分别表示常温和高温条件下, 减数分裂间期的行为特征; B和H分别表示常温和高温条件下, 减数分裂前期I-细线期的行为特征; C和I分别表示常温和高温条件下, 减数分裂前期I-偶线期的行为特征; D和J分别表示常温和高温条件下, 减数分裂前期I-粗线期的行为特征; E和K分别表示常温和高温条件下, 减数分裂前期I-双线期的行为特征; F和L分别表示常温和高温条件下, 四分体时期的行为特征。NT: 常温; HT: 高温。标尺=20 μm。"

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