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Photosynthetic Properties and Gene Cloing of Rice (Oryza Sativa) Albino Mutants albi and alblhc2

Author: LiJiaFei
Tutor: MiHuaLing
School: Shanghai Normal University
Course: Botany
Keywords: Albino Mutant Photosynthetic characteristics Gene mapping Functional studies Protein interactions Rice
CLC: S511
Type: Master's thesis
Year: 2011
Downloads: 60
Quote: 2
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Abstract

Chloroplasts of higher plants, photosynthesis spaces, their normal development depends on the coordinated expression of nuclear genes and chloroplast genes, and mutations in these genes lead to defects in chloroplast development and photosynthetic characteristics change. In our work, respectively, of two rice Albino the mutant albl and alblhc2 the photosynthetic characteristics were studied, and the mutant gene map-based cloning and functional analysis of the results are as follows: 1 Rice albino to mutation the body albl gene positioning by X-ray mutagenesis and functional studies, we get a case of Rice albino mutant albl performance of the mutant seedlings period for the first piece completely leaves albino incomplete leaf, light green leaves all albino about a month and death . The genetic analysis showed that the albino trait caused by a single recessive nuclear gene mutation. Chlorophyll fluorescence analysis showed that, the PS Ⅱ under normal conditions in a climatic chamber mutant seedlings grown for 2 weeks and PS Ⅰ activity decreased significantly. The chloroplast ultrastructure display mutant light green part of the chloroplast thylakoid membrane structure arranged irregularly, almost non-existent in the albino leaf chloroplast stroma thylakoid membranes. The results showed that the the mutant albl protein - pigment complex accumulation has decreased by gentle blue-green gel electrophoresis separation of photosynthetic pigments complex. The thylakoid membrane Western blot analysis showed that mutant chloroplast gene encoding the core protein of the photosynthetic system has significantly reduced. The use of molecular markers, we InDel use map-based cloning isolated gene ALBL. The sequencing showed that the mutant albl gene deletion of five bases CCATG, wherein the ATG start codon of the gene coding sequence in the base mutation causes the gene not normally encoded target protein. Genomic DNA of Agrobacterium-mediated genetic complementation experiments, homozygous mutant plants revert to a normal phenotype. We named the gene ALBL (ALBINO LETHALITY), the analysis found that the gene encodes a DYW-type PPR proteins in chloroplasts, sequence analysis showed that the protein contains 11 PPR domain, an E domain, an E domain a the DYW structure domain. Immunoprecipitation method, we the four candidate chloroplast protein interactions with ALBL, yeast two-hybrid experiments confirmed that one of the the Os03g0214900 gene encoding the protein SRI protein interactions with the PPR. Between specific sites of action, and the study of the interaction mechanism in continuing. Rice albino mutation the body alblhc2 photosynthetic characteristics and gene mapping by 60Coγ-ray mutagenesis, we get an example of Rice Albino Mutant alblhc2 of the mutant seedlings during the incomplete leaf is green, completely leaves the majority of white blades gradually All albino died, until a month or so. The genetic analysis showed that the trait is controlled by a single recessive nuclear gene. LHC Ⅱ trisomy in the mutant significant degradation of PS II core protein and pigment protein significantly reduced maximal photochemical efficiency reduce serious disruption of electron transport, chlorophyll fluorescence parameters Fo significantly increased. Using molecular markers of InDel, the gene is located on chromosome 4 the mark Os40719 and Os408 between a range of about 1,535 kb. These results eventually cloning and study of the role of the gene in rice chloroplast development has laid a foundation.

Photosynthetic Properties and Gene Cloing of Rice (Oryza Sativa) Albino Mutants albi and alblhc2

Abstract

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