论文总字数：17999字

摘 要

敌草隆是一种内吸传导型取代脲类除草剂。过量的敌草隆残留会对植物的生理机能会产生影响，导致靶标或非靶标植物枯黄，叶片枯萎，甚至产生灭生作用。水稻作为主要的农作物之一，其安全生产关乎民生大计，开展水稻降解农药机制研究具有很重要的现实意义。MiR156是一个植物中重要的microRNA，调控植物许多重要的生理发育过程。近期研究发现MiR156调控植物对非生物胁迫，如：重金属。然而在农药降解方面还研究较少。因此，本文根据MiR156在水稻中的表达量，制定相关实验，就MiR156调控下OsSPL介导的水稻代谢敌草隆的机制初探。本论文选择两种野生型水稻株系（日本晴和秀水）、micro156过表达株系（MiR156OE）以及micro156缺失突变株系（Mimc156-3）为实验材料，敌草隆为处理药剂。研究敌草隆处理对四种株系的生长情况的影响，结果发现MiR156OE未加药时长势要优于其对照组日本晴，但在添加敌草隆后，MiR156OE的生物量相较于其对应野生型株系（日本晴）大幅下滑，其差距被明显缩小，无显著差异；Mimc156-3与其野生型株系（秀水）在加药与未加药处理下的生长情况无明显差异。进一步研究了敌草隆在四种株系的积累情况。结果发现，MiR156OE与日本晴水稻株内敌草隆含量无明显差异，Mimc156-3和秀水的敌草隆积累情况与前者类似。可见，MiR156介导的OsSPL基因与水稻响应和代谢敌草隆相关性不高，相关实验结果为后续OsSPL调控作物对非生物胁迫耐受调控机制提供理论参考。

关键词：水稻； 敌草隆； MiR156； 积累量

STUDY ON THE EFFECT OF MIR156-DEPENDENT OSSPL ON THE BIOTRANSFORMATION OF DIURON IN RICE

Abstract

Diuron is a new type of herbicide, which can replace urea herbicide. Excessive residues of diuron will affect the physiological functions of plants, leading to the target or non target plants wither, leaves wither, and even have a killing effect.

As one of the main crops, the safety of rice production is related to people's livelihood. It is of great practical significance to study the mechanism of rice degradation pesticides. MiR156 is an important microRNA in plants, which regulates many important physiological development processes. Recent studies have found that mir156 regulates abiotic stress in plants, such as heavy metals. However, there are few researches on pesticide degradation. Therefore, based on the expression of MiR156 in rice, this paper made relevant experiments to explore the mechanism of rice metabolism of diuron mediated by OSSPL under the regulation of MiR156.

In this paper, two wild-type rice lines (Qinghe Xiushui, Japan), MiR156OE and Mimc156-3 were selected as experimental materials, and diuron was used as treatment agent. The results showed that the growth of MiR156OE was better than that of control group, but the biomass of MiR156OE decreased significantly compared with that of its corresponding wild-type strain (Nippon Qing) after adding diuron, and the difference was significantly reduced without significant difference; the growth of Mimc156-3 and its wild-type strain (Xiushui) was increased with and without adding diuron There was no significant difference in the growth of the treated plants. The accumulation of diuron in four strains was further studied. The results showed that there was no significant difference in diuron content between MiR156OE and Nippon clear rice. The accumulation of diuron in Mimc156-3 and Xiushui was similar to the former. It can be seen that the correlation between rice response and metabolism of diuron and osspl gene mediated by MiR156 is not high. The relevant experimental results provide a theoretical reference for the subsequent regulation mechanism of OSPL on abiotic stress tolerance of crops.

Key words: Rice; Diuron; MiR156; Accumulation;

目录

摘 要 I

Abstract II

第一章 文献综述 1

1.1 前言 1

1.2水稻简介 1

1.3敌草隆简介 2

1.3.1 敌草隆的作用机理与使用现状 2

1.4 MiR156与SPL 3

1.5 植物中农药的检测方法 4

1.5.1 毛细管电泳法（CE） 4

1.5.2 超高效液相色谱法 4

1.5.3 气相色谱法 4

第二章 实验材料及方法 5

2.1 实验材料 5

2.1.1 供试材料 5

2.1.2 实验材料 5

2.2 试验仪器 6

2.3 实验方法 6

2.3.1 对水稻种子的发芽处理以及对幼苗的培养 6

2.3.2 敌草隆的制备 8

2.3.3 水稻的加药培养以及对照试验 8

2.3.4 各位置的敌草隆提取方法与液相条件 8

第三章 实验结果与讨论 11

3.1 敌草隆对不同基因型水稻的生长的影响 11

3.1.1 不同品种水稻幼苗地上部分的生长状态 11

3.1.2 不同水稻根长叶长数据 13

3.2 敌草隆对不同基因型水稻的叶绿素的影响 15

3.3不同基因型水稻的积累敌草隆情况 16

3.3.1 标准曲线的绘制与求得检出限 16

3.3.2 水稻叶中敌草隆含量数据 16

3.3.3 水稻根中敌草隆含量数据 17

3.3.4 营养液中敌草隆含量数据 18

3.4 讨论与分析 19

3.4.1 MiR156与SPL基因的能力 19

3.4.2 敌草隆可能对水稻产生的影响 20

3.4.3 思考MiR156调控下SPL介导的水稻代谢敌草隆的机制与实验结论 20

第四章 分析与展望 22

参考文献 23

致 谢 25

第一章 文献综述

1.1 前言

中国作为一个拥有庞大人口基数的国家，日常的饮食起居相较于其他国家而言要面对更为严峻的考验。而水稻作为一种家喻户晓的粮食作物，在解决人民温饱问题中扮演着举足轻重的角色。全世界大约有一半的人口把稻米当作主食。根据预测，随着世界人口的不断增加，到2030年水稻产量至少需要增加40%才能满足世界庞大人口对于其需求。在水稻的生长过程中只有严格管理，防治病虫害才能保障人们对于粮食作物的需求。为了获取更高产量，更优品质的水稻，我们往往会使用农药，达到除虫、除草、抗菌的效果。

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