线粒体呼吸在水杨酸信号调控毛果杨对黑斑病抗性中的作用毕业论文

 2021-04-20 11:04

摘 要

为了初步解释毛果杨的抗病分子基础,探究线粒体呼吸电子传递在水杨酸信号调控毛果杨对黑斑病抗性中的作用,本实验利用毛果杨和杨生褐盘二孢菌为实验材料,进行毛果杨-杨生褐盘二孢菌互作研究。实验使用实时荧光定量PCR等技术,对毛果杨在应答侵染过程中的呼吸途径相关基因以及水杨酸途径相关基因的表达做定量分析。主要研究结果如下:

(1)杨生褐盘二孢菌接种毛果杨离体叶片,导致叶片只出现褐色斑点,无显著黑斑病症状。

(2)杨生褐盘二孢菌接种后导致SA合成相关基因PAL1ICS在24hpi上调表达,信号转导相关基因PR1在48hpi被激活。

(3)杨生褐盘二孢菌接种后导致JA合成相关基因AOC1在24hpi下调表达,JA信号抑制子编码基因JAZ10a在24和48hpi均显著上调表达,JA下游核心转录因子MYC2基因表达在24和48hpi均显著受到抑制;同时,ET合成相关的ACO3和ET受体编码基因ETR1表达量在24hpi均极剧烈下调,EIN3基因表达变化不显著。

(4)杨生褐盘二孢菌接种后导致线粒体呼吸电子传递链复合体I、III和IV(COX6b)编码基因24hpi强烈下调,交替氧化酶基因AOX1b;48hpi时,Mn SOD表达增强而UCP表达未上调,甚至被抑制。

关键词:毛果杨;杨生褐盘二孢菌;线粒体呼吸链;水杨酸途径

The Role of Mitochondrial Respiration in Salicylic Acid Signal Regulation of Populus trichocarpa Resistance to Black Spot Disease

ABSTRACT

To explore the role of mitochondrial electron transport chain in salicylic acid (SA) signaling in the the resistance of Populus trichocarpa to Marssonina brunnea f. sp. multigermtubi, The interaction between P. trichocarpa and M. brunnea was studied. The expression of genes involved in mitochondrial electron transport chain, as well as the biosynthesis and signaling of SA, jasmonic acid (JA) and ethylene (ET) in vitro leaf of P. trichocarpa were analyzed by real-time quantitative PCR . The main findings are as follows:

(1) The in vitro leaves of Populus trichocarpa were inoculation with M. brunnea f. sp. Multigermtubi, which led to only brown spots in the leaves and no significant symptoms of black spot disease.

(2) Genes PAL1 and ICS involved in SA synthesis were up-regulated at 24hpi, and PR1 mediating were signaling activated in 48hpi.

(3) The expression of JA synthesis related gene AOC1 was down-regulated in 24hpi, the expression of JA signal inhibitor coding gene JAZ10a was significantly up-regulated in 24 and 48hpi, and the expression of core transcription factor MYC2 gene downstream of JA was significantly inhibited in 24 and 48hpi.At the same time, the expression of genes ACO3 related to ET synthesis and ET receptor coding genes ETR1 was significantly down-regulated in 24hpi, but the expression of EIN3 gene did not change significantly.

(4) Genes encoding the complex I (NUDO), complex III (UCRH) and IV (COX6b) in mitochondrial electron transport chain was strongly down-regulated at 24hpi. The expression of Mn SOD increased but the expression of UCP was inhibited while AOX1b encoding alternative oxidase was inoculated 48hpi.

Key words: Populus trichocarpa; M. brunnea f. sp. multigermtubi; miETH; SA pathway

目 录

1 前言 1

1.1实验研究背景 1

1.1.1杨树的形态特征及研究价值 1

1.1.2杨树黑斑病及研究杨树黑斑病的意义 1

1.1.3植物激素在生物胁迫响应中的作用 2

1.1.4 线粒体呼吸与生物胁迫 3

1.2实验目的及研究意义 4

2 材料与方法 5

2.1实验材料 5

2.1.1毛果杨叶片 5

2.1.2杨生褐盘二孢菌菌株 5

2.1.3试剂盒 5

2.2实验方法 5

2.2.1毛果杨叶片的获取 5

2.2.2杨生褐盘二孢菌菌株的接种 6

2.2.3RNA提取及反转录 6

2.3统计学分析 9

3 结果与分析 10

3.1实验现象 10

3.2 水杨酸途径相关基因表达量的变化 10

3.3茉莉酸/乙烯途径相关基因表达量的变化 11

3.4线粒体呼吸链相关基因表达量的变化 13

4 结论 17

致谢 18

参考文献 19

1 前言

1.1实验研究背景

1.1.1杨树的形态特征及研究价值

杨树是被子植物门(Angiosperms),双子叶植物纲(Dicots)杨柳科(Salicaceae)杨属(Populus)的落叶乔木。杨树广泛分布于我国华中、华北、西北、东北等广阔地区,它对自然条件的适应性极强,是优秀的工业材用树种,也是优秀的防护林树种,生长速度也极为迅速,因此成为我国重要的材用树种,在我国大部分地区种植有大量的杨树人工林。

毛果杨(Populus trichocarpa)是从美国引进的品种,属杨属青杨派,具有易繁殖、生长迅速的特点。2006年它的基因组序列已被测出并公布,因此被作为木本转基因植物中的模式植物,是林木树种遗传转化研究中的典型代表种。毛果杨主要分布在北美洲西部地区,树形挺拔,枝繁叶茂,生长高度可达30米以上。在中国由于受毛果杨材料难以得到、枝条扦插不易成活的限制,用组织培养技术可以获得一定量的毛果杨苗木。

1.1.2杨树黑斑病及研究杨树黑斑病的意义

杨树黑斑病是侵害杨树幼树和苗木的主要病害之一。杨树黑斑病,又名杨树褐斑病,是一种真菌型病害,能够侵染多种杨树,在我国杨树分布区均有发生[1]。病害的表现为叶片上出现斑点,初期为针刺状的红色发亮斑点,继续扩大为黑色圆斑,在中间出现乳白色分生孢子堆后,病斑开始向黑褐色转变,当大量黑斑连成片,即导致叶片枯死、掉落,严重时甚至会使整棵树木发生早衰,使杨树的年生长量和年平均木材产量受到严重损失。

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