摘 要

杨树是世界上适应性最强，分布最广的树种之一，是我国重要的造林绿化、工业用材树种，但是杨树的病害的问题始终制约着杨树的高效生产。杨盘二孢菌作为杨树黑斑病的主要致病菌，可以引起叶片黑斑并降低杨树光合效率，最终导致木材产量降低。本选题以南林895为材料，研究杨树-杨生褐盘二孢菌互作中茉莉酸、乙烯和水杨酸相关基因的响应，以及呼吸作用相关基因的变化。结果如下

1. 杨生褐盘二孢菌接种南林895杨离体叶片后，叶片出现黑斑病症状。
2. 杨生褐盘二孢菌接种后导致JA和ET合成相关基因在24hpi上调表达，信号转导相关基因48hpi被激活；但对SA合成基因影响不大，信号转导相关基因PR1和SA结合蛋白编码基因被抑制。
3. 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和AOX编码基因上调，但复合体III和IV编码基因下调； UCP2表达增强而MnSOD表达被抑制。

关键词：杨树；杨生褐盘二孢菌；乙烯；茉莉酸；线粒体呼吸电子传递

The role of mitochondrial electron transport chain in the regulation of jasmonic acid/ethylene signaling in the ‘Nanlin 895’- Marssonina brunnea interaction

ABSTRACT

Populus is one of the most adaptable and widely distributed tree species in the world. It is an important afforestation and industrial timber tree species in China .However, the problem of populus diseases always restricts the efficient production of poplar .M. brunnea f. sp. Multigermtubi as the main pathogen of Poplar black spot disease, It can cause black spots in leaves and reduce photosynthetic efficiency of poplar, which can eventually lead to a decrease in wood output.‘Nanlin 895’is the material of this paper .The expression of genes involved in mitochondrial electron transport chain, as well as the biosynthesis and signaling of salicylic acid（SA）, jasmonic acid (JA) and ethylene (ET) in vitro leaf of ‘Nanlin 895’ were analyzed by real-time quantitative PCR . The main findings are as follows:

(1)The in vitro leaves of ‘Nanlin 895’ were inoculation with M. brunnea f. sp. Multigermtubi, which led to black spot on the leaves.

(2)Gene related to jasmonic acid (JA) and ethylene (ET) biosynthesis were up-regulated at 24hpi,and mediating were signaling activated in 48hpi. However, it had little influence on gene related to salicylic acid（SA）biosynthesis, PR1 mediating and KGDHE2 were signaling restrained in 48hpi.

(3)Genes encoding the complex I (NUDO) and AOX in mitochondrial electron transport chain was up-regulated but complex III (UCRH) and complex IV(COX6b) was down-regulated. The expression of UCP2 increased but the expression of MnSOD was inhibited 48hpi.

Key words：Populus；M. brunnea f. sp. Multigermtubi；Ehylene；Jsmonic acid；Mitochondrial electron transport chain

目 录

1 前言………………………………………………………………………………………1

1.1 实验研究背景………………………………………………………………1

1.1.1 杨树的形态特征和研究价值……………………………………1

1.1.1 研究杨树黑斑病的意义………………………………………………1

1.2 目前研究进展…………………………………………………………………………1

1.2.1 杨树-杨生褐盘二孢菌互作的研究方向……………………………1

1.22 杨树-杨生褐盘二孢菌互作的研究进展及成果……………………………2

1.3 实验研究方法…………………………………………………………………………3

1.3.1 植物RNA的提取方法……………………………………………3

1.3.2 实时荧光定量PCR技术……………………………………………3

2 材料与方法………………………………………………………………………………5

2.1 实验材料……………………………………………………………………5

2.1.1 杨树叶片…………………………………………………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 植株和病原菌准备及病原菌接种……………………………………5

2.2.2 RNA提取及反转录…………………………………………………6

2.3 统计学分析…………………………………………………………………………8

3 实验结果………………………………………………………………………………9

3.1 发病表现……………………………………………………………………9

3.2 茉莉酸/乙烯相关基因表达量的变化……………………………………………9

3.3 水杨酸相水杨酸关基因表达量的变化…………………………………………11

3.3 呼吸相关基因表达量的变化………………………………………………………13

结论 ………………………………………………………………16

致谢 …………………………………………………………………17

参考文献 …………………………………………………………………18

1 前言

1.1 实验研究背景

1.1.1 杨树的形态特征和研究价值

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。^[1]

南林895速生杨：雌株，是美洲黑杨×欧美杨F1无性系，具有美洲黑杨的基本形态特征。乔木，树形高大，干形

通直圆满，尖削度小，分枝粗度中等，树皮薄；叶片大，心形，叶绿色，淡绿色；1年生枝条颜色青绿色，枝条基部形状为圆形，中上部微呈五边形，木质化棱线明显；皮孔中下部为圆形，上为椭圆形，皮孔大小为2-6毫米；芽长卵形，浅紫红色，基部绿色，长6-8毫米，顶端尖，不紧贴（梢部芽）。南林895杨喜光、喜水、喜肥，在光照水肥充足的情况下，生长十分迅速。抗溃疡病及褐斑病，较耐干旱瘠薄，具有较强的无性繁殖能力和遗传稳定性^[2-5]。

由南京林业大学杨树研究开发中心在“九五”期间选出的杨树单板用材新品种。其性状与南林95杨相同，生长量略大于南林95杨，育苗和造林成活率略低于南林95杨，为杨树抗性的研究提供了基础。