本文選題：水稻　切入點：黃葉突變體　出處：《湖南農(nóng)業(yè)大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

【摘要】：作物的光能利用率與作物的產(chǎn)量息息相關(guān),而水稻葉色突變體是研究水稻光合作用的理想材料。本研究以超級稻品種中嘉早17(YK17)的黃葉突變體yl1為研究材料,對突變體yl1進行表型鑒定與精細定位,主要研究結(jié)果如下：1)性狀調(diào)查結(jié)果表明：yl1 在全生育期內(nèi)莖、葉和穗都表現(xiàn)為淡黃色,同時株高降低、每穗粒數(shù)減少、劍葉寬變窄和粒長變長,其他性狀如分蘗數(shù)、穗長、結(jié)實率、千粒重、劍葉長和粒厚則無顯著變化,yl1 花粉育性正常。2)葉綠素含量測定結(jié)果表明：苗期YK17葉片中葉綠素a、葉綠素b及總?cè)~綠素含量都高于yl1 ,差異極顯著,其中葉綠素b含量為yl1的6倍,總?cè)~綠素含量為yl1的2.5倍。抽穗期YK17的葉片中葉綠素b含量高于yl1,是yl1的3倍,差異極顯著：而葉綠素a含量略有升高,但總的葉綠素含量依舊極顯著低于YK17,為YK17的二分之一。抽穗期yl1的穗中的葉綠素a、葉綠素b及總?cè)~綠素含量都顯著低于YK17,含量僅為YK17的一半,因此穗也表現(xiàn)為淡黃色。同時由葉綠素a與葉綠素b的比值分析得知,葉片中葉綠素含量的下降主要是由于葉綠素b的下降,而穗中葉綠素含量的降低則主要是由于葉綠素a和葉綠素b的同步下降。光合特性分析表明：yl1的凈光合速率和氣孔導(dǎo)度與YK17相比都有所降低。葉綠體的電鏡觀察表明：yl1葉肉細胞內(nèi)的葉綠體數(shù)目明顯低于YK17。3)遺傳分析結(jié)果表明：yl1基因為單隱性核基因,通過圖位克隆將該基因定位在Chrl短臂中端44.8kb內(nèi),預(yù)測區(qū)間內(nèi)有4個開放閱讀框(ORFs),其中ORF1(Os01g17170)編碼鎂原卟啉Ⅸ單甲脂環(huán)化酶,測序分析發(fā)現(xiàn)該基因CDS序列的第515個堿基由C變?yōu)門,從而導(dǎo)致編碼蛋白的第172個氨基酸由絲氨酸變?yōu)楸奖彼�。RT-PCR實驗表明ORF1在突變體的葉和穗中的表達量都低于野生型,由此確認Os01g17170為yl1的候選基因。亞細胞定位實驗將YL1蛋白定位在細胞質(zhì)內(nèi)。
[Abstract]:The light energy utilization efficiency of crops is closely related to the yield of crops, and the rice leaf color mutant is an ideal material for studying photosynthesis. In this study, the yellow leaf mutant yl1 of Zhongjia Zongzao YK17) was used as the research material. Phenotypic identification and fine mapping of the mutant yl1 were carried out. The main results were as follows: 1) the results showed that the stem, leaves and panicles of the mutant were yellowish in the whole growth period, and the plant height was decreased, and the number of grains per panicle was decreased. Other traits such as tiller number, ear length, seed setting rate, 1000-grain weight, There was no significant change in leaf length and grain thickness. The results showed that chlorophyll a, chlorophyll b and total chlorophyll content in YK17 leaves at seedling stage were higher than those in yl1, and the difference was very significant. The content of chlorophyll b was 6 times of that of yl1, and the content of total chlorophyll was 2.5 times of that of yl1. The content of chlorophyll b in leaves of YK17 at heading stage was 3 times higher than that of yl1, and the content of chlorophyll a was slightly higher than that of yl1. However, the total chlorophyll content of yl1 was still significantly lower than that of YK17, which was 1/2 of that of YK17. The contents of chlorophyll a, chlorophyll b and total chlorophyll in panicle of yl1 at heading stage were all significantly lower than that of YK17, and the content was only half of that of YK17. From the analysis of the ratio of chlorophyll a to chlorophyll b, it was found that the decrease of chlorophyll content in leaves was mainly due to the decrease of chlorophyll b. The decrease of chlorophyll content in ear was mainly due to the simultaneous decrease of chlorophyll a and chlorophyll b. The analysis of photosynthetic characteristics showed that the net photosynthetic rate and stomatal conductance of 1 / 1 were lower than those of YK17. The results showed that the number of chloroplasts in the mesophyll cells was significantly lower than that in YK17.3). The gene was located within 44.8 kb of Chrl short arm by map-cloning, and four open reading frames were predicted in the region, in which ORF1 + Os01g17170) encodes magnesium protoporphyrin IX monomethylenecyclic enzyme. Sequencing analysis showed that the 515th base of the CDS sequence of the gene changed from C to T, which led to the transformation of the 172nd amino acid of the encoded protein from serine to phenylalanine. RT-PCR results showed that the expression of ORF1 in the leaves and panicles of the mutant was lower than that in the wild type. Therefore, Os01g17170 was identified as a candidate gene for yl1. Subcellular localization of YL1 protein in cytoplasm was performed.
【學(xué)位授予單位】：湖南農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S511;Q943.2

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