Proteome Analysis and Morphological Characteristics of Purple Pericarp Rice

Mohammad Mominur Rahman,Mohammad Nurul Matin,Saifullah Ahmed Saief,Sang Gu Kang 한국작물학회 2010 한국작물학회 학술발표대회 논문집 Vol.2010 No.04

Rice with purple colored pericarp deposit anthocyanin on the seed coat and color accumulation increased rapidly during seed development. The purple color of rice pericarp is genetically determined by the Prp locus. Inheritance of purple pericarp was studied in Prp/ Kumgangbyeo (indica type Korean variety). Pericarp color of the F1 plants was purple and the F2 population of 274 plants segregated into 3 purple: 1 white ratio indicating dominant nature of the purple color. Comparative proteomic approaches using 2-DE were applied to analyze the protein profiles and molecular mechanism of purple color formation in ricepericarp. Results revealed that approximately 1,500protein spots were reproducibly detected in the gels with silver staining across the two biological replicates. Among them, 46 proteins were expressed differentially between purple color pericarp rice and white color pericarp of the wild type rice, in which 28 and 16 protein spots were more than two fold up regulated in the wild type and purple pericarp, respectively. MALDI-TOF MS analysis of nine spots revealed that putative fructokinase,embryo-specific protein and one unknown proteins were abundant in the wild type, whereas, anthocyanidin synthase, putative chloroplast inner envelope protein, and dihydroflavonol reductase were highly abundant in the Prp rice. Results indicated anthocyanidin synthase and/or dihydroflavonol reductase might be involved in the biosynthetic pathway of the purple color formation in the rice pericarp. [This research was supported by the Grant funded by Agricultural R&D Promotion Center, ARPC (IPET project number: 108091-05-1-CG000)].

Comparative Phenotypic and Physiological Characteristics of Spotted Leaf 6 (spl6) and Brown Leaf Spot2 (bl2) Lesion Mimic Mutants (LMM) in Rice

Mohammad Nurul Matin,Saifullah Ahmed Saief,Mohammad Mominur Rahman,Dong Hoon Lee,강호덕,Dong Sun Lee,강상구 한국분자세포생물학회 2010 Molecules and cells Vol.30 No.6

Spontaneous necrotic lesions were found in a lesion mimic mutant brown leaf spot 2 (bl2) without pathogenic infection. Small spots in the seedlings appeared at the four leaves stage and gradually grew into a large round and black area with a gray center on the leaf surfaces. Lower growth habit and lower agronomic trait values with reduced stature, tiller, and panicle number, as well as lower yield potential were noted in the mutants relative to the trait values of the wild-type plants. Microscopic analysis revealed that mesophyll chloroplast was severely damaged or absent in the spotted area of the mutant leaves. Total chlorophyll content, hydrogen peroxide level, and catalase activity were increased at up to 45 days after germination and were dropped at 60 d in the mutant leaves. However, the total protein contents were reduced slightly with a growth period of up to 45 days and were increased at 60 days after germination. A gradual increment of the total ascorbic acid contents in the mutants were observed with advanced plant age, but increased until 45 days and dropped comparatively at 60 days in the wild-type leaves. Increased gene transcrip-tions of OsPDI and OsGPX1 were noted in the spotted leaves as compared to the non-spotted leaves of the mutant and wild-type leaves, where-as transcripts of OsTPX were transcribed at lower levels in the spotted leaves as compared to the non-spotted leaves. The genetic nature of the bl2 mutant indicated that the F1 plants evidenced the wild-type phenotype and that bl2 was governed by a single recessive gene.

The Genetic Constitutions of Complementary Genes Pp and Pb Determine the Purple Color Variation in Pericarps with Cyanidin-3-O-glucoside Depositions in Black Rice

Md Mominur Rahman,이경은,이은선,Mohammad Nurul Matin,이동선,윤정식,김종배,강상구 한국식물학회 2013 Journal of Plant Biology Vol.56 No.1

The purple pericarp color in rice was controlledby two dominant complementary genes, Pb and Pp. Crossingblack rice ‘Heugnambyeo’ variants with three varieties of whitepericarp rice gave a segregation ratio of 9 purple: 3 brown:4 white. The Pp genes were segregated by homozygousPpPp alleles for the dark purple pericarps, heterozygousPppp alleles for the medium and mixed purple pericarps, andhomozygous pppp alleles for either brown or white pericarpswith a 1 PpPp: 2 Pppp: 1 pppp segregation ratio, indicatingthat the Pp allele in rice is incompletely dominant to therecessive pp allele. Among the purple seeds, the amount ofcyanidin-3-O-glucoside was higher in the dark purple seeds(Pb_PpPp) than in the medium purple seeds (Pb_Pppp). Moreover, no cyanidin-3-glucoside was detected in brown(Pb_pppp) or white pericarp seeds (pbpbpppp). Thesefindings indicated that the level of cyanidin-3-glucoside wasdetermined by the copy number of the Pp allele. Furthergenotype investigation of the F3 progeny demonstrated thatthe dominant Pb allele was present in either purple or brownpericarp. A 2-bp (GT) deletion from the DNA sequences ofthe dominant and functional Pb was found in the same DNAsequences of the recessive and non-functional pb allele. These findings suggested that the presence of at least adominant Pb allele was an essential factor for colordevelopment in rice pericarps. In conclusion, the Pp allele inrice is incompletely dominant to the recessive pp allele; thus,the number of dominant Pp alleles determines the concentrationof cyanidin-3-O-glucoside in black rice.

Comparative Phenotypic and Physiological Characteristics of Spotted Leaf 6 (spl6) and Brown Leaf Spot2 (bl2) Lesion Mimic Mutants (LMM) in Rice

Matin, Mohammad Nurul,Saief, Saifullah Ahmed,Rahman, Mohammad Mominur,Lee, Dong-Hoon,Kang, Ho-Duck,Lee, Dong-Sun,Kang, Sang-Gu Korean Society for Molecular and Cellular Biology 2010 Molecules and cells Vol.30 No.6

Spontaneous necrotic lesions were found in a lesion mimic mutant brown leaf spot 2 (bl2) without pathogenic infection. Small spots in the seedlings appeared at the four leaves stage and gradually grew into a large round and black area with a gray center on the leaf surfaces. Lower growth habit and lower agronomic trait values with reduced stature, tiller, and panicle number, as well as lower yield potential were noted in the mutants relative to the trait values of the wild-type plants. Microscopic analysis revealed that mesophyll chloroplast was severely damaged or absent in the spotted area of the mutant leaves. Total chlorophyll content, hydrogen peroxide level, and catalase activity were increased at up to 45 days after germination and were dropped at 60 d in the mutant leaves. However, the total protein contents were reduced slightly with a growth period of up to 45 days and were increased at 60 days after germination. A gradual increment of the total ascorbic acid contents in the mutants were observed with advanced plant age, but increased until 45 days and dropped comparatively at 60 days in the wild-type leaves. Increased gene transcriptions of OsPDI and OsGPX1 were noted in the spotted leaves as compared to the nons-potted leaves of the mutant and wild-type leaves, whereas transcripts of OsTPX were transcribed at lower levels in the spotted leaves as compared to the non-spotted leaves. The genetic nature of the bl2 mutant indicated that the $F_1$ plants evidenced the wild-type phenotype and that bl2 was governed by a single recessive gene.

Comparative Characteristics of Two Lesion Mimic Mutants, Brown Leaf Spot 2 (bl2) and Spotted Leaf 6 (spl6) in Rice

Saifullah Ahmed Saief,Mohammad Nurul Matin,Mohammad Mominur Rahman,Sang Gu Kang 한국작물학회 2010 한국작물학회 학술발표대회 논문집 Vol.2010 No.04

A comparative phenotypic study between bl2 and spl6 mutant wasperformed to characterize spot formation mechanisms of bl2 mutant. Small spots appeared at the seedling stage in bl2 and later it covered large round areas on the leaves whereas, relatively small red spots in parallel line on both leaf surfaces at late tillering stage were observed in spl6. Vegetative and reproductive growth was reduced due to lesion formation at early age in the mutants. Lower growth habit and agronomic trait value was observed in mutants as compared to wild type plants. Genetic segregation data among F2 population revealed that both mutants are recessive in nature. Mesophyll chloroplast was not found in spotted area which demonstrates the damage of chloroplast cell at spotted area due to cell death. Transmission electron microscopy also confirmed the chloroplast damage. Increased level of total chlorophyll and hydrogen peroxide content were observed till 45 days of growth after transplantation under natural environment and dropped at 60 days. Catalase activity was increased until 45 days and decreased at 60 days whereas very slight level difference in protein content was observed. However, increasing level of total ascorbic acid contents were found in spl6 and bl2 as compared with wild type till 60 days after transplantation. Higher expressions of OsPDI and OsGPX1 in bl2 spotted leaves were found whereas OsTPX expression was very low in the spotted leaf. (This research was supported by the National Research foundation of Korea, Grant 0070065).