水稻DA1家族基因调控水稻粒型的功能研究

发布时间：2024-04-11 02:04

　　籽粒大小是决定水稻产量的主要因素之一。籽粒的长度和宽度与粒重的呈正相关,改良粒型对提高水稻产量具有十分重要的作用。作为重要的农艺性状,粒型是由多基因控制的数量性状,遗传基础比较复杂,易受到环境影响。目前一些控制水稻粒长、粒宽和粒重的基因已被克隆,但控制粒型的分子和遗传调控网络仍不清楚。拟南芥DA1基因编码一个泛素受体蛋白,能够通过调控细胞分裂影响拟南芥器官和种子大小。DA1包含两个LIM结构域,分别位于位于N端和C端。DA1通过LIM结构域与泛素相互作用。拟南芥中有7个DA1的同源基因,其编码蛋白被命名为DA1相关(DAR)蛋白。在水稻中DA1的同源基因有4个分别命名为OsDA1、OsDAR2、OsDAR3和OsDAR4。本论文通过对水稻DA1相关(DAR)基因的研究表明:OsDAR2、OsDAR3和OsDAR4可以调节水稻粒宽和粒长,增加水稻的千粒重和单株产量。本研究表明过表达OsDAR2、OsDAR3和OsDAR4基因可以导致水稻粒宽和粒长同时增加从而导致千粒重和单株总粒重增加,而CAS9敲除株系则表现完全相反的表型,籽粒的长度和宽度同时减小进而千粒重和单株总粒重同时减少。这表明O...

【文章页数】：81 页

【学位级别】：博士

【文章目录】：
摘要
abstract
CHAPTER 1 INTRODUCTION
    1.1 Rice, an important cereal crop
    1.2 Rice a model plant for genetics and breeding aspects
    1.3 Seed formation mechanism and final seed size
        1.3.1 Zygotic and maternal factors regulate organ size in plants
    1.4 Superior grain quality with high grain yield
        1.4.1 Superior grain quality
        1.4.2 High grain yield
    1.5 VP16 and EAR
        1.5.1 Domain structure and interacting proteins of VP
        1.5.2 EAR transcription suppression domain
    1.6 DA1 and DAR Genes(Research Progress)
        1.6.1 Control of final seed and organ size by the DA1 gene family in Arabidopsisthaliana
        1.6.2 The ubiquitin receptor DA1 interacts with the E3 ubiquitin ligase DA2 toregulate seed and organ size in Arabidopsis
        1.6.3 Control of root meristem size by DA2 in Arabidopsis
        1.6.4 The ubiquitin receptor DA1 regulates seed and organ size by modulating thestability of the ubiquitin-specific protease UBP15/SOD2 in Arabidopsis
        1.6.5 The ubiquitin receptors DA1, DAR1, and DAR2 redundantly regulateendoreduplication by modulating the stability of TCP14/15
        1.6.6 Genetic and molecular network for maternal control of seed size
CHAPTER 2 MATERIALS AND METHODS:
    2.1 PCR primers
        2.1.1 Tips for a good primer design
    2.2 DNA extraction
        2.2.1 DNA extraction by CTAB method
    2.3 RNA extraction
    2.4 c DNA synthesis
    2.5 Full length c DNA amplification
    2.6 Gel electrophoresis
        2.6.1 Agarose gel electrophoresis
        2.6.2 SDS-PAGE
    2.7 Western blotting
    2.8 DNA agarose gel recovery
    2.9 E. coli transformation
        2.9.1 Heat-shock transformation
        2.9.2 Single colony culture
        2.9.3 PCR assay
    2.10 Plasmid extraction
    2.11 Agrobacterium electroporation
    2.12 Gateway reactions
    2.13 Chemical preparation
CHAPTER 3 RESULTS
    3.1 DAR genes in Arabidopsis and Rice
        3.1.1 LOC_Os03g16090
        3.1.2 LOC_Os12g40490
        3.1.3 LOC_Os03g42820
    3.2 DARs-VP16 transgenic lines
        3.2.1 Expression level of DARs- VP16 in transgenic lines
        3.2.2 DARs fused with VP16 enhances grain size in rice
        3.2.3 DARs-VP16 lines showed compact panicle and more spikelets
        3.2.4 DARs-VP16 lines showed no obvious architecture phenotype
    3.3 DARs-EAR transgenic lines
        3.3.1 Expression level of DARs-EAR in transgenic lines
        3.3.2 DARs fused with EAR reduces grain size and KGW in rice
        3.3.3 DARs fused with EAR reduces number of spikelets per panicle fused
        3.3.4 DARs-EAR lines showed no obvious architecture phenotype
    3.4 DARs Over expression (DAR-OEX) lines
        3.4.1 Expression level of DARs in DARs-OEX lines
        3.4.2 DARs-OEX enhances grain length and width
        3.4.3 DARs-OEX lines showed more spikelets with normal seed setting rate
        3.4.4 DAR-OEX lines shows no significant change in plant height
    3.5 DAR4-CAS9 knock out lines
        3.5.1 Knockout of DAR4 reduces grain length and width in rice
        3.5.2 DAR4 knockout lines reduces number of spikelets per panicles
        3.5.3 Plant architecture of knockout lines was similar with wild type
    3.6 Expression level of DARs in rice plant tissues at different stages
CHAPTER 4 DISCUSSION
CHAPTER 5 CONCLUSIONS
REFERENCE
Acknowledgement
Resume



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