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Genetic analysis and gene fine mapping of a rolling leaf mutant (rl_(11(t))) in rice (Oryza sativa L.)

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Chinese Science Bulletin 2010年第17期55卷 1763-1769页

作者： ZHOU Yong FANG YunXia ZHU JinYan LI ShengQiang gu Fei gu minghong LIANG guoHua Jiangsu Key Laboratory for Crop Genetics and Physiology/Jiangsu Key Laboratory of Plant Functional Genomics Yangzhou University Yangzhou 225009 China

The exploration of new genes controlling rice leaf shape is an important foundation for rice functional genomics and plant architecture improvement. In the present study, we identified a rolling leaf mutant from indica variety Yuefeng B, named rl11(t), which exhibited reduced plant height, rolling and narrow leaves. Leaves in rl11(t) mutant showed abnormal number and morphology of veins compared with those in wild type plants. In addition, rl11(t) mutant was less sensitive to the inhibitory effect of auxin than the wild type. Genetic analysis suggested that the mutant was controlled by a single recessive gene. Gene Rl11(t) was initially mapped between SSR markers RM6089 and RM124 on chromosome 4. Thirty-two new STS markers around the Rl11(t) region were developed for fine mapping. A physical map encompassing the Rl11(t) locus was constructed and the target gene was finally delimited to a 31.6 kb window between STS4-25 and STS4-26 on BAC AL606645. This provides useful information for cloning of Rl11(t) gene.

关键词：功能基因组学精细定位遗传分析突变体 Oryza 水稻动叶叶片形状

Microdissection and amplification of the chromosome arm 5S in a rice telo-tetrasomic

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Chinese Science Bulletin 1998年第7期43卷 590-594页

作者： Zhukuan Cheng Huihuanq Yan Benyuan Dang Zanmin Hu minghong gu Lihuang Zhu Chinese Acad Sci Inst Genet Beijing 100101 Peoples R China Yangzhou Univ Coll Agr Yangzhou 225009 Peoples R China

The rice aneuploids with telochromosomes are ideal genetic stocks for chromosome arm identification and microdissection. A rice telo-tetrasomic line with two extra short arms of chromosome 5 (5S) was used in the present study. The arms of 5S were microdissected from the prometaphase cells in mitosis and amplified with linker adaptor PCR (LA-PCR). The amplified fragments, ranging from 200-3000 bp, were confirmed to be from the rice chromosome arm 5S by the rice STS and microsatellite markers.

关键词： telo-tetrasomic chromosome arm microdissection rice.

Genetic analysis and gene fine mapping for a rice novel mutant(rl_(9(t)))with rolling leaf character

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Chinese Science Bulletin 2006年第1期51卷 63-69页

作者： YAN Changjie YAN Song ZHANG Zhengqiu LIANG guohua LU Jufei gu minghong Jiangsu Key Laboratory of Plant Functional Genomics/Jiangsu Key Laboratory for Crop Genetics and Physiology Yangzhou University Yangzhou 225009 China

Leaf shape is an important parameter for ideotype breeding in rice, and the rolling of leaf is also beneficial to efficient ripening of grains. This encourages the explorations of new genes that regulate leaf shape. In this study, genetic analysis and gene mapping were carried out for a novel rolling leaf mutant identified from japonica variety Zhonghua 11. The SSR marker analysis showed that the mutant was controlled by a single recessive gene (rl9(t)) lo- cated on chromosome 9. Fine mapping of the Rl9(t) locus was conducted with 30 new STS markers de- veloped around Rl9(t) anchored region based on the sequence diversity between Nipponbare and 93-11. The fine mapping necessitated the contruction of a PAC contig encompassing the Rl9(t) locus, which was delimited to a 42 kb region. This could therefore en- hance the cloning of the target gene in further stud- ies.

关键词：水稻遗传分析分子标记基因工程

Development and characterization of interspecific hybrids between Oryza sativa and O. latifolia by in situ hybridization

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Chinese Science Bulletin 2008年第19期53卷 2973-2980页

作者： YI ChuanDeng TANG ShuZhu ZHOU Yong LIANG guoHua GONG ZhiYun gu minghong Jiangsu Province Key Laboraty of Crop Genetics and Physiology/Key Laboraty of Plant Functional Genomics of Ministry of Education Yangzhou University Yangzhou 225009 China

Oryza sativa and O. latifolia belong to the AA and CCDD genomes of Oryza, respectively. In this study, interspecific hybrids of these species were obtained using the embryo rescue technique. Hybrid panicle traits, such as long awns, small grain, exoteric large purple stigma, grain shattering and dispersed panicles, resemble that of the paternal parent, O. latifolia, whereas there is obvious heterosis in such respects as plant height, tillering ability and vegetative vigor. Chromosome pairing and the genomic components of the hybrid were subsequently investigated using genomic in situ hybridization (GISH) and fluorescent in situ hybridization (FISH) analysis. Based on the mitotic metaphase chromosome numbers of the root tips investigated, the hybrid is a triploid with 36 chromosomes. The genomic constitution of the hybrid is ACD. In the meiotic metaphase I of the hybrid pollen mother cell, poor chro- mosome pairing was identified and most of the chromosomes were univalent, which resulted in com- plete male sterility in the hybrid.

关键词：漂白亚麻纤维卷胚芽种间杂交技术

Generating of rice OsCENH3-GFP transgenic plants and their genetic applications

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Chinese Science Bulletin 2008年第19期53卷 2981-2988页

作者： YU HengXiu WANG Xin GONG ZhiYun TANG Ding gu minghong CHENG ZhuKuan Key Laboratory of Crop Genetics and Physiology of Jiangsu Province/Key Laboratory of Plant Functional Genomics of Ministry of Education Yangzhou University Yangzhou 225009 China Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing 100101 China

In order to investigate rice functional centromeres, OsCENH3-GFP chimeric gene was constructed and transformed into the indica rice variety, Zhongxian 3037, mediated by Agrobacturium. The integration of the exogenous genes in the transgenic plants was confirmed by PCR and Southern blotting. The transgenic plants grow normally during their whole life time, just like Zhongxian 3037. No significant defects were detected in either mitosis or meiosis of the transgenic plants. The overlapping of GFP signals and anti-CENH3 foci in both mitotic and meiotic cells from T0 and T1 generation plants indicated that GFP had been successfully fused with CENH3, so the GFP signals can well represent the CENH3 locations on each chromosome. To evaluate the applicability of the transgenic plants to other genetic studies, fluorescence in situ hybridization (FISH) using rice centromeric tandem repetitive sequence CentO as the probe was conducted on the zygotene chromosomes of pollen mother cells (PMCs). It has been revealed that the GFP signals are overlapping with CentO FISH signals, showing that CentO is one of the key elements constituting rice functional centromeres. Immunofluorescent staining using anti-α-tublin antibody and anti-PAIR2 antibody on the chromosomes during mitosis and meiosis stages of the transgenic plants further reveals that OsCENH3-GFP transgenic plants can be widely used for studying rice molecular biology, especially for tagging functional centromeres in both living cells and tissues.

关键词：着丝点稻子遗传物质栽培技术

Introduction of antisense waxy gene into main parent lines of indica hybrid rice

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Chinese Science Bulletin 2002年第14期47卷 1192-1197页

作者： CHEN Xiuhua LIU Qiaoquan WANG Zongyang WANG Xingwen CAI Xiuling ZHANG Jingliu gu minghong Key Laboratory of Biotechnology of Jiangsu Province Yangzhou UniversityYangzhou 225009China Shanghai Institute of Plant Physiology and Ecology Chinese Academy of ScienceShanghai 200032China

Amylose content in rice endosperm is one of the key determinants of rice eating and cooking quality, and the poor quality of indica hybrid rice is closely related to the high amylose level in rice grains. In order to improve the grain quality of the indica hybrid rice by genetic engineering, an antisense fragment of rice waxy gene, driven by the 5’-franking sequences of the rice waxy gene, was successfully introduced into three major parent lines of indica hybrid rice, all contain a high amylose level in the grains, via Agrobacte-rium, and more than 100 hygromycin-resistant plants were regenerated. The analysis of PCR amplification and Southern blots indicated that the T-DNA containing the antisense waxy gene had been integrated into the genome of transgenic rice plants. Most of the primary transgenic rice plants grew normally, and the mature seeds from these transgenic plants were performed for analysis of the amylose content. The results showed that the amylose content in the endosperm of some grains

关键词： transgenic rice indica rice maintain line antisense waxy gene amylose content.

Molecular mapping of two semidwarf genes in an indica rice variety Aitaiyin3(Oryza sativa L.)

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Frontiers in Biology 2006年第2期1卷 93-98页

作者： Zhao Xiangqiang Liang guohua Zhou Jingsong Yan Changjie Cao Xiaoying gu minghong Key Laboratory of Plant Function Genomics the Ministry of EducationAgricultural CollegeYangzhou UnivercityYangzhou 225009China

Genetic analysis established that Aitaiyin3,a dwarf rice variety derived from a semidwarf cultivar Taiyin1,carries two recessive semidwarf *** using simple sequence repeat(SSR)markers,we mapped the two semidwarf genes,sd-1 and sd-t2 on chromosomes 1 and 4,***-t2 was thus named because the semidrawf gene sd-t has already been identified from Aitaiyin 2 whose origin could be traced back to *** result of the molecular mapping of sd-1 gene revealed it is linked to four SSR markers found on chromosome *** markers are:RM297,RM302,RM212,and OSR3 spaced at 4.7 cM,0 cM,0.8cM and 0 cM,***-t2 was found to be located on chromosome 4 using five SSR markers:two markers,SSR332 and RM1305 located proximal to sd-t2 are spaced 11.6 cM,3.8 cM,respectively,while the three distally located primers,RM5633,RM307,and RM401 are separated by distances of 0.4 cM,0.0 cM,and 0.4 cM,respectively.

关键词： rice semidwarf gene SSR markers molecular mapping

Isolation and Identification of a Functional Centromere Element in the Wild Rice Species Oryza granulata with the GG Genome

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Journal of Genetics and Genomics 2015年第12期42卷 699-702页

作者： Chuandeng Yi Mingsen Wang Wei Jiang Derong Wang Yong Zhou Zhiyun Gong guohua Liang minghong gu Jiangsu Key Laboratory of Crop Genetics and Physiology Co-Innovation Center for Modern Production Technology of Grain Crops Key Laboratory of Plant Functional Genomics of the Ministry of Education Yangzhou University

The centromere of eukaryotic chromosomes is the crucial locus responsible for sister chromatid cohesion and for correct segregation of chromosomes to daughter cells during cell division. In the structural genomics era, centromeres represent the last frontiers of higher eukaryotic genomes because of their densely methylated, highly repetitive and, heterochromatic DNA （Hall et al., 2004）. Although these functions are conserved among all eukaryotes, centromeric DNA sequences are evolving rapidly （Jiang et al., 2003）.

关键词： FISH BAC Isolation and Identification of a Functional Centromere Element in the Wild Rice Species Oryza granulata with the GG Genome

Genetic analysis and molecular mapping of a presenescing leaf gene psll in rice （Oryza sativa L.）

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Chinese Science Bulletin 2006年第24期51卷 2986-2992页

作者： WANG dun WU Shujun ZHOU Yong ZHOU Lihui XU Jiefen HU Jing FANG Yunxia gu minghong LIANG guohua Key Laboratory of the Ministry of Education for Plant Functional Genomics/Jiangsu Key Laboratory for Crop Genetics and Physiology Yangzhou University Yangzhou 225009 China

A rice psl1 (presenescing leaf) mutant was obtained from a japonica variety Zhonghua 11 via radiation of 60Co-γ in M2 generation. Every leaf of the mutant began to wither after it reached the big-gest length,while the leaves of the wild variety could keep green for 25―35 d. In this study,genetic analysis and gene mapping were carried out for the mutant identified. The SSR marker analysis showed that the mutant was controlled by a single recessive gene (psl1) located on chromosome 2. Fine mapping of the psl1 locus was conducted with 34 new STS markers developed around psl1 anchored region based on the sequence diversity between Nippon-bare and 93-11. The psl1 was further mapped be-tween two STS markers,STS2-19 and STS2-26,with genetic distances of 0.43 and 0.11 cM,respectively,while cosegregated with STS2-25. A BAC contig was found to span the psl1 locus,the region being delim-ited to 48 kb. This result was very useful for cloning of the psl1 gene.

关键词： psIl基因水稻叶片早衰遗传分析分子图谱