Characterization of Phenotypes of Rice Lesion Resembling Disease Mutants

Mohammad Nurul Matin,Hak Soo Suh,Sang Gu Kang 한국유전학회 2006 Genes & Genomics Vol.28 No.3

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.

Genetic and Phenotypic Analysis of lax1-6, a Mutant Allele of LAX PANICLE1 in Rice

Mohammad Nurul Matin,Sang Gu Kang 한국식물학회 2012 Journal of Plant Biology Vol.55 No.1

Proper function of the LAX1 gene is required for the development of axillary meristem in rice. Here, we report genetic and phenotypic characters of a novel recessive mutant allele of rice LAX1 gene, lax1-6, which showed abnormal panicle phenotypes with few numbers of elongated primary rachis branches. Beside typical lax mutant phenotype, abnormalities of lax1-6 mutant allele were observed with defect lemma and palea primordial in floral organs. The lax1-6 mutant locus was linked between SSR markers RM7594 and RM5389 on chromosome 1 with 1.02% and 1.0% recombination frequencies, respectively. Molecular analysis revealed that the lax1-6 mutant allele was caused by a transversion mutation of nucleotide T to G substitution that resulted in an amino acid substitution from serine (S) to alanine (A) at the 117th position from amino terminus of a basic helix-loop-helix protein coded by LAX1gene. Furthermore, we found that the Oryza sativa indica type cv. IRRI347 contained 24 nucleotide deletion in the upstream sequence in the LAX1 gene, but this deletion did not influence panicle morphology, which demonstrated that the deletion is a polymorphism in rice. All together, the lax1-6 mutant is a newly identified allele of LAX1 gene displaying the abnormal axillary meristems and inflorescences in rice.

Phenotypic and Genotypic Analysis of Rice Lesion Mimic Mutants

Matin, Mohammad Nurul,Pandeya, Devendra,Baek, Kwnag-Hyun,Lee, Dong-Sun,Lee, Jai-Heon,Kang, Ho-Duck,Kang, Sang-Gu The Korean Society of Plant Pathology 2010 Plant Pathology Journal Vol.26 No.2

Lesion mimic mutant (LMM) plants display spontaneous necrotic lesions on their leaves without any pathogenic infection. Specific rice LMMs designated as spotted leaf (spl) including spl1, spl3, spl4, spl5 and spl6 are genetically known as lesion resembling disease (lrd) mutant. The inheritance patterns in the $F_1$ and $F_2$ progenies of these mutants are controlled by recessive genetic factors. Lesion development in the rice LMMs were controlled by both development stages and environmental factors. The rice LMMs exhibited higher numbers of spots under $45^{\circ}C$ temperature than those under $30^{\circ}C$. Contents of chlorophyll were drastically reduced at 60 days old LMM leaves when the spot formation was severe. The levels of endogenous hydrogen peroxide were highest at 45 days old mutants but reduced at 60 days old. Transcription levels of stress related genes including thioredoxin peroxidase and protein disulfide isomerase were reduced in spotted leaves than those of non spotted leaves. It could be suggested that scavenging system against reactive oxygen species induced by either stresses or innate metabolisms may not work properly in the rice LMMs. As these rice LMMs autonomously expressed clear lesions of lrd phenotype without pathogen infection, it could be useful to understand stresses responses in plants.

Phenotypic and Genotypic Analysis of Rice Lesion Mimic Mutants

Mohammad Nurul Matin,Devendra Pandeya,백광현,이동선,이재현,강호덕,강상구 한국식물병리학회 2010 Plant Pathology Journal Vol.26 No.2

Lesion mimic mutant (LMM) plants display spontaneous necrotic lesions on their leaves without any pathogenic infection. Specific rice LMMs designated as spotted leaf (spl) including spl1, spl3, spl4, spl5 and spl6 are genetically known as lesion resembling disease (lrd)mutant. The inheritance patterns in the F1 and F2progenies of these mutants are controlled by recessive genetic factors. Lesion development in the rice LMMs were controlled by both development stages and environmental factors. The rice LMMs exhibited higher numbers of spots under 45oC temperature than those under 30oC. Contents of chlorophyll were drastically reduced at 60 days old LMM leaves when the spot formation was severe. The levels of endogenous hydrogen peroxide were highest at 45 days old mutants but reduced at 60days old. Transcription levels of stress related genes including thioredoxin peroxidase and protein disulfide isomerase were reduced in spotted leaves than those of non spotted leaves. It could be suggested that scavenging system against reactive oxygen species induced by either stresses or innate metabolisms may not work properly in the rice LMMs. As these rice LMMs autonomously expressed clear lesions of lrd phenotype without pathogen infection, it could be useful to understand stresses responses in plants.

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.

Genetic Characterization of Lax Panicle (Lax) Mutant and Linkage Analysis with Spotted leaf 6 (spl6) Mutant in Rice

Mohammad Nurul Matin,Sang Gu Kang 한국작물학회 2010 한국작물학회 학술발표대회 논문집 Vol.2010 No.04

Rice lax panicle (lax) mutant had defect in early inflorescence differentiation and lateral branch formation and exhibitedfewer numbers of rachis branches and spikelets in the panicle. The mutant also had abnormality in floral organ formation. Whereas, the spotted leaf 6 (spl6) mutant produced lesions in absence of pathogen. Analysis with a lax spl6 double mutant indicated both genes are controlled by single recessive factor and linked on chromosome 1 with 7 centiMorgan map distance. The lax gene was fine mapped within the 261.3 kb region between RM7594 and RM5389 that predicted 10 open reading frames (ORF) as candidate for lax gene. Sequencing of the candidates in the lax mutant revealed a substitution of nucleotide T by G corresponding to an amino acid substitution from serine (S) to alanine (A) at the 117th position within the coding region of the candidate ORF bhlh123. The intronless 1080 bp cDNA of LAX gene contains an ORF of 215 amino acids and encoded a basic helix-loop-helix transcription factor. Analysis of a 5.1 kb genomic fragment of the lax gene from homozygous dominant progeny which corresponded to indica cultivar resulted in a long deletion of 24 nucleotides in the upstream of LAX cDNA. (This research was supported by the National Research foundation of Korea, Grant 0070065).

Phenotypic, Genetic and Proteomic Characterization of Lesion Mimic Mutants in Rice

Mohammad Nurul Matin,Sang Gu Kang 한국작물학회 2008 한국작물학회 학술발표대회 논문집 Vol.2008 No.10

We have discussed here the phenotypic and genetic characteristics as well as proteomic analysis of lesion mimic mutants (LLM) in rice. LLM is one of the mutants that induces cell death without infection of pathogen and produces defense signaling pathways. As the phenotypic expression, most LLMs induce spots on the leaf blades and leaf sheaths at their various developmental stages. We have discussed the nature of bl1, bl2, spl1, spl3, spl4, spl5 and spl6 LLMs in rice which were formed developmentally controlled spot on the leaf blades that were appeared as tiny dotted spots during tillering stage and gradually increased up to maturity. Through Northern blot analysis lower levels of rubisco large subunit and rubisco small subunit were observed in spotted leaves (sp) compared to non-spotted leaves (nsp). However, catalase was severely degraded in the sp. Broken thylakoid membranes of mesophyll chloroplasts were seen in nsp sections and were absent in sp sections of the mutant. Through 2-DE analysis 159 protein spots were differentially expressed between wild type and mutant from identified 800 reproducibleproteins, where 114 spots were up-regulated and 45 were down-regulated. Among quantified 25 protein spots, except two, all of the protein spots including protein disulfide isomerase, transketolase, thioredoxin peroxidase, ATP synthase, and rubisco large and small subunits were identified in the wild type but were absent in the mutant. However, catalase was up-regulated in the mutant. Genetic analysis indicated that studied bl1, bl2 and spl6 mutants are controlled by a single recessive gene.

Morphological Characteristics of the Rice (Oryza sativa L.) with Red Pigmentation

Mohammad Nurul Matin(모하마드누룰마틴),Sang Gu Kang(강상구) 한국생명과학회 2010 생명과학회지 Vol.20 No.1

벼의 적색 종피 표현형질은 Rc 혹은 Rd 그리고 RdRc 두 개의 복수유전자에 의하여 결정된다. 본 연구에서는 종피에 색소가 있는 돌연변이 벼 7계통을 벼 표현형 돌연변이 군들로부터 분리하였다. 이들 종피색의 돌연변이 형질을 조사한 결과 두 개의 변이체가 갈색의 종피형질을 가졌으며 네 개의 변이체가 적색의 종피형질을 가졌다. 그리고 한 개의 변이체는 종피에 색깔이 없었다. 수확 후 종자의 종피색 표현형질에 관한 정밀조사를 실시한 결과 두개의 갈색종피 돌변이체는 Rc 유전자형 이었으며 네개의 적색종피 돌변이체는 RdRc 유전자형이었다. 또한 종피에 색깔이 없는 한 개의 돌변이체가 Rd 유전자형이었다. 이들 중 동일한 종피색깔을 나타내는 돌연변이 개체들이 단일 돌연변이 개체의 후손인지 여부를 판명하기 위하여 돌변이체들의 초형과 농업형질을 조사한 결과 동일 종피색 돌변이체들의 형질이 서로 상이하므로 각각 서로 다른 돌연변이 계통들로 확인되었다. 그리고 이들 종피색깔 돌연변이들은 초형, 분얼 특징, 임성과 수량성 등을 포함한 농업형질들이 정상 벼의 농업형질들보다 대체로 빈약하고 또한 높은 탈립성을 보였다. 이러한 종피색과 관련된 돌연변이 벼 유전자원은 안토시아닌과 폴리페놀이 풍부하므로 병충해 저항성 품종과 환경스트레스 내성 품종의 육종에 필요할 뿐만 아니라 종피의 색깔을 결정하는 유전자들의 기능을 밝히는 데 중요한 자료로 사용될 것이다. Seven rice germplasms with red pigmentation within the pericarp were isolated from a large mutant collection. These red pericarp phenotypes resulted from the functions of the Rc, Rd and RdRc genes. Among them, two brown pericarp of the Rc type, four red pericarp of the RdRc type, and one white pericarp of the Rd type were identified. Morphological and agronomic characteristics of those rice germplasms were studied. The Rc type germplasms have the faint red or brown color pericarp and the Rd types produce the white pericarp, whereas the RdRc type germplasms have the dark red pericarp. Most of the important agronomic characteristics including plant stature, tillering ability, spikelet fertility, and total grain yield were lower in the colored rice than those of the wild-type control. All of the studied colored rice germplasms had a tendency of easy seed-shattering in comparison to the control. These characteristics of newly identified germplasms will be useful for identifying the genes responsible for pericarp color phenotype determination.

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)].