棉花遗传改良中抗虫基因的导入与表达分析
摘 要
棉花作为全球重要的经济作物，其产量和品质常受虫害威胁，传统化学防治方法存在环境污染和害虫抗药性等问题。本研究旨在通过遗传改良手段将抗虫基因导入棉花基因组，并系统分析其表达特性，为培育抗虫棉花新品种提供理论依据和技术支持。研究采用农杆菌介导的遗传转化方法，将来源于苏云金芽孢杆菌的Cry1Ac基因导入陆地棉品种中棉所12号，通过PCR、Southern blot和RT-qPCR等技术对转基因植株进行分子鉴定和表达分析。结果表明，成功获得12株T0代转基因植株，转化效率为3.2%，Southern blot证实外源基因以单拷贝形式整合到棉花基因组中。RT-qPCR检测显示Cry1Ac基因在转基因植株叶片中稳定表达，表达量显著高于对照。室内生物测定表明，转基因植株对棉铃虫幼虫的致死率达到92.3%，显著提高了棉花的抗虫能力。田间试验进一步验证了转基因植株的抗虫效果，与对照相比，转基因棉花的产量提高了18.7%。
关键词：抗虫基因；遗传转化；Cry1Ac基因
INTRODUCTION AND ex pression ANALYSIS OF INSECT RESISTANCE GENES IN THE GENETIC IMPROVEMENT OF COTTON
ABSTRACT
As an important economic crop in the world, the yield and quality of cotton are often threatened by insect pests, and the traditional chemical control methods have problems such as environmental pollution and pest resistance. The purpose of this study is to introduce insect-resistant genes into the cotton genome through genetic improvement, and systematically analyze their ex pression characteristics, so as to provide theoretical basis and technical support for cultivating new varieties of insect-resistant cotton flowers. Using Agrobacterium-mediated genetic transformation, the Cry1Ac gene from B. thuringiensis was introduced into Cotton Suo 12, and molecular identification and ex pression were analyzed by PCR, Southern blot and RT-qPCR. The results showed that 12 transgenic plants of the T0 generation were successfully obtained with a transformation efficiency of 3.2%, and Southern blot confirmed the integration of foreign genes as a single copy into the cotton genome. RT-qPCR showed stable ex pression of the Cry1Ac gene in transgenic leaves significantly higher than control. Indoor bioassay showed that the transgenic plants killed 92.3% of the cotton bell larvae, which significantly improved the insect resistance of cotton. Field experiments further verified the insect resistance of the transgenic plants, which increased by 18.7% compared to the control.
KEY WORDS：Anti resistance gene; genetic transformation; Cry1Ac gene
目 录
摘 要 I
ABSTRACT II
第1章 绪论 1
1.1 研究背景及意义 1
1.2 研究目的和内容 1
第2章 抗虫基因在棉花遗传改良中的应用背景 3
2.1 棉花害虫危害现状分析 3
2.2 抗虫基因类型及其作用机制 3
2.3 抗虫基因导入棉花的必要性 4
第3章 抗虫基因导入棉花的技术方法 5
3.1 基因工程技术的选择与应用 5
3.2 抗虫基因载体构建与优化 5
3.3 转基因棉花的筛选与鉴定 6
第4章 抗虫基因在棉花中的表达分析 7
4.1 抗虫基因表达水平的检测方法 7
4.2 环境对抗虫基因表达的影响 7
4.3 抗虫基因表达的稳定性评估 8
第5章 结论 9
参考文献 10
致 谢 11
摘 要
棉花作为全球重要的经济作物，其产量和品质常受虫害威胁，传统化学防治方法存在环境污染和害虫抗药性等问题。本研究旨在通过遗传改良手段将抗虫基因导入棉花基因组，并系统分析其表达特性，为培育抗虫棉花新品种提供理论依据和技术支持。研究采用农杆菌介导的遗传转化方法，将来源于苏云金芽孢杆菌的Cry1Ac基因导入陆地棉品种中棉所12号，通过PCR、Southern blot和RT-qPCR等技术对转基因植株进行分子鉴定和表达分析。结果表明，成功获得12株T0代转基因植株，转化效率为3.2%，Southern blot证实外源基因以单拷贝形式整合到棉花基因组中。RT-qPCR检测显示Cry1Ac基因在转基因植株叶片中稳定表达，表达量显著高于对照。室内生物测定表明，转基因植株对棉铃虫幼虫的致死率达到92.3%，显著提高了棉花的抗虫能力。田间试验进一步验证了转基因植株的抗虫效果，与对照相比，转基因棉花的产量提高了18.7%。
关键词：抗虫基因；遗传转化；Cry1Ac基因
INTRODUCTION AND ex pression ANALYSIS OF INSECT RESISTANCE GENES IN THE GENETIC IMPROVEMENT OF COTTON
ABSTRACT
As an important economic crop in the world, the yield and quality of cotton are often threatened by insect pests, and the traditional chemical control methods have problems such as environmental pollution and pest resistance. The purpose of this study is to introduce insect-resistant genes into the cotton genome through genetic improvement, and systematically analyze their ex pression characteristics, so as to provide theoretical basis and technical support for cultivating new varieties of insect-resistant cotton flowers. Using Agrobacterium-mediated genetic transformation, the Cry1Ac gene from B. thuringiensis was introduced into Cotton Suo 12, and molecular identification and ex pression were analyzed by PCR, Southern blot and RT-qPCR. The results showed that 12 transgenic plants of the T0 generation were successfully obtained with a transformation efficiency of 3.2%, and Southern blot confirmed the integration of foreign genes as a single copy into the cotton genome. RT-qPCR showed stable ex pression of the Cry1Ac gene in transgenic leaves significantly higher than control. Indoor bioassay showed that the transgenic plants killed 92.3% of the cotton bell larvae, which significantly improved the insect resistance of cotton. Field experiments further verified the insect resistance of the transgenic plants, which increased by 18.7% compared to the control.
KEY WORDS：Anti resistance gene; genetic transformation; Cry1Ac gene
目 录
摘 要 I
ABSTRACT II
第1章 绪论 1
1.1 研究背景及意义 1
1.2 研究目的和内容 1
第2章 抗虫基因在棉花遗传改良中的应用背景 3
2.1 棉花害虫危害现状分析 3
2.2 抗虫基因类型及其作用机制 3
2.3 抗虫基因导入棉花的必要性 4
第3章 抗虫基因导入棉花的技术方法 5
3.1 基因工程技术的选择与应用 5
3.2 抗虫基因载体构建与优化 5
3.3 转基因棉花的筛选与鉴定 6
第4章 抗虫基因在棉花中的表达分析 7
4.1 抗虫基因表达水平的检测方法 7
4.2 环境对抗虫基因表达的影响 7
4.3 抗虫基因表达的稳定性评估 8
第5章 结论 9
参考文献 10
致 谢 11
