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1. SUPER WOMAN 2 (SPW2) 通过抑制花同源异型基因OsMADS3OsMADS58OsMADS13DROOPING LEAF的表达维持小穗的器官特征
ZHUANG Hui, LAN Jin-song, YANG Qiu-ni, ZHAO Xiao-yu, LI Yu-huan, ZHI Jing-ya, SHEN Ya-lin, HE Guang-hua, LI Yun-feng
Journal of Integrative Agriculture    2024, 23 (1): 59-76.   DOI: 10.1016/j.jia.2023.07.010
摘要287)      PDF    收藏

水稻的花器官特征主要由ABCE四类基因决定,它们大多编码MADS-box转录因子。然而,在花发育过程中,对于这些基因的表达如何被调控的研究很少。本研究报道了一个名为SUPER WOMAN 2SPW2)的基因,该基因在水稻的小穗/小花发育过程中通过调控雌蕊特征基因OsMADS3OsMADS13OsMADS58DL的表达发挥重要作用。SPW2突变导致小穗内的护颖、外稃、内稃、浆片和雄蕊中出现异位的柱头/子房状组织。通过图位克隆,我们揭示了SPW2编码一个植物特有的类EMF1蛋白,该蛋白是PRC2复合物的重要组成部分,并介导H3K27me3修饰。表达分析显示,SPW2突变导致OsMADS3OsMADS13OsMADS58DL在小穗的非雌蕊器官中异位表达。此外,ChIP-qPCR结果显示这些基因在染色质上的H3K27me3修饰水平显著降低。因此,我们的研究结果表明SPW2通过参与H3K27me3介导的雌蕊特征基因表观遗传沉默,进而调控它们在水稻小穗的非雌蕊器官中的表达。这项研究拓宽了我们对于SPW2通过表观遗传调控花器官特征基因的分子机制的认识。

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2. Identification and gene mapping of the starch accumulation and premature leaf senescence mutant ossac4 in rice
ZHU Mao-di, CHEN Xin-long, ZHU Xiao-yan, XING Ya-di, DU Dan, ZHANG Ying-ying, LIU Ming-ming, ZHANG Qiu-li, LU Xin, PENG Sha-sha, HE Guang-hua, ZHANG Tian-quan
Journal of Integrative Agriculture    2020, 19 (9): 2150-2164.   DOI: 10.1016/S2095-3119(19)62814-5
摘要140)      PDF    收藏
The rice mutant ossac4 (starch accumulating 4) was raised from seeds of the rice (Oryza sativa L.) indica maintainer line Xinong 1B treated with ethyl methanesulfonate.  The distal and medial portions of the second leaf displayed premature senescence in the ossac4 mutant at the four-leaf stage.  Physiological and biochemical analysis, and cytological examination revealed that the ossac4 mutant exhibited the premature leaf senescence phenotype.  At the four-leaf stage, the leaves of the ossac4 mutant exhibited significantly increased contents of starch compared with those of the wild type (WT).  Quantitative real-time PCR analysis showed that the expression levels of photosynthesis-associated genes were down-regulated and the expression levels of glucose metabolism-associated genes were abnormal.  Genetic analysis indicated that the ossac4 mutation was controlled by a single recessive nuclear gene.  The OsSAC4 gene was localized to a 322.7-kb interval between the simple-sequence repeat marker XYH11-90 and the single-nucleotide polymorphism marker SNP5300 on chromosome 11.  The target interval contained 20 annotated genes.  The present results demonstrated that ossac4 represents a novel starch accumulation and premature leaf senescence mutant, and lays the foundation for cloning and functional analysis of OsSAC4.
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3. Identification of long-grain chromosome segment substitution line Z744 and QTL analysis for agronomic traits in rice
MA Fu-ying, DU Jie, WANG Da-chuan, WANG Hui, ZHAO Bing-bing, HE Guang-hua, YANG Zheng-lin, ZHANG Ting, WU Ren-hong, ZHAO Fang-ming
Journal of Integrative Agriculture    2020, 19 (5): 1163-1169.   DOI: 10.1016/S2095-3119(19)62751-6
摘要118)      PDF    收藏
Length of grain affects the appearance, quality, and yield of rice.  A rice long-grain chromosome segment substitution line Z744, with Nipponbare as the recipient parent and Xihui 18 as the donor parent, was identified.  Z744 contains a total of six substitution segments distributed on chromosomes (Chrs.) 1, 2, 6, 7, and 12, with an average substitution length of 2.72 Mb.  The grain length, ratio of length to width, and 1 000-grain weight of Z744 were significantly higher than those in Nipponbare.  The plant height, panicle number, and seed-set ratio in Z744 were significantly lower than those in Nipponbare, but they were still 78.7 cm, 13.5 per plant, and 86.49%, respectively.  Furthermore, eight QTLs of different traits were identified in the secondary F2 population, constructed by Nipponbare and Z744 hybridization.  The grain weight of Z744 was controlled by two synergistic QTLs (qGWT1 and qGWT7) and two subtractive QTLs (qGWT2 and qGWT6), respectively.  The increase in the grain weight of Z744 was caused mainly by the increase in grain length.  Two QTLs were detected, qGL1 and qGL7-3, which accounted for 25.54 and 15.58% of phenotypic variation, respectively.  A Chi-square test showed that the long-grain number and the short-grain number were in accordance with the 3:1 separation ratio, which indicates that the long grain is dominant over the short-grain and Z744 was controlled mainly by the principal effect qGL1.  These results offered a good basis for further fine mapping of qGL1 and further dissection of other QTLs into single-segment substitution lines.
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4. Gene mapping and candidate gene analysis of aberrant-floral spikelet 1 (afs1) in rice (Oryza sativa L.)
ZHANG Ting, YOU Jing, YU Guo-ling, ZHANG Yi, CHEN Huan, LI Yi-dan, YE Li, YAO Wan-yue, TU Yu-jie, LING Ying-hua, HE Guang-hua, LI Yun-feng
Journal of Integrative Agriculture    2020, 19 (4): 921-930.   DOI: 10.1016/S2095-3119(19)62847-9
摘要136)      PDF    收藏
The spikelet is a unique inflorescence structure in grasses.  However, the molecular mechanism that regulates its development remains unclear, and we therefore characterize a spikelet mutant of rice (Oryza sativa L.), aberrant-floral spikelet 1 (afs1), which was derived from treatment of Xinong 1B with ethyl methanesulfonate.  In the afs1 mutant, the spikelet developed an additional lemma-like organ alongside the other normally developed floral organs, and the paleae were degenerated to differing degrees with or without normally developed inner floral organs.  Genetic analysis revealed that the afs1 phenotype was controlled by a single recessive gene.  The AFS1 gene was mapped between the insertion/deletion (InDel) marker Indel19 and the simple sequence repeat marker RM16893, with a physical distance of 128.5 kb on chromosome 4.  Using sequence analysis, we identified the deletion of a 5-bp fragment and a transversion from G to A within LOC_Os04g32510/ LAX2, which caused early termination of translation in the afs1 mutant.  These findings suggest that AFS1 may be a new allele of LAX2, and is involved in the development of floral organs by regulating the expression of genes related to their development.  The above results provide a new view on the function of LAX2, which may also regulate the development of spikelets.
 
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5. Identification and QTL mapping of Z550, a rice backcrossed inbred line with increased grains per panicle
WANG Shi-ming, CUI Guo-qing, WANG Hui, MA Fu-ying, XIA Sai-sai, LI Yun-feng, YANG Zheng-lin, LING Ying-hua, ZHANG Chang-wei, HE Guang-hua, ZHAO Fang-ming
Journal of Integrative Agriculture    2019, 18 (3): 526-531.   DOI: 10.1016/S2095-3119(18)61996-3
摘要253)      PDF(pc) (310KB)(469)    收藏
An elite backcrossed inbred line Z550 with increased grains per panicle was identified from advanced backcrosses between Nipponbare and Xihui 18 by simple sequence repeat (SSR) marker-assisted selection (MAS).  Z550 carries 13 substitution segments distributed on chromosomes 1, 6, 7, 8, 9, 10, and 12, with an average substitution length of 1.68 Mb.  Compared with the Nipponbare parental line, plant height, panicle length, spikelets per panicle, grains per panicle, and grain weight for Z550 were significantly increased.  While the grain width of Z550 was significantly narrower, and the seed setting ratio (81.43%) was significantly lower than that of Nipponbare, it is still sufficient for breeding purposes.  Quantitative trait loci (QTLs) mapping for important agronomic traits was conducted with the F2 population derived from Nipponbare crossed with Z550 using the restricted maximum likelihood (REML) method.  A total of 16, including 12 previously unreported QTLs were detected, with contribution rates ranging from 1.46 to 10.49%.  Grains per panicle was controlled by 8 QTLs, 5 of which increased number of grains whereas 3 decreased it.  qGPP-1, with the largest contribution (10.49%), was estimated to increase grains per panicle by 30.67, while qGPP-9, with the minimum contribution rate (2.47%), had an effect of increasing grains per panicle by 15.79.  These results will be useful for further development of single segment substitution lines with major QTLs, and for research of their molecular functions via QTL cloning.
 
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6. Gene mapping and candidate gene analysis of multi-floret spikelet 3 (mfs3) in rice (Oryza sativa L.)
ZHENG Hao, ZHANG Jun, ZHUANG Hui, ZENG Xiao-qin, TANG Jun, WANG Hong-lei, CHEN Huan, LI Yan, LING Ying-hua, HE Guang-hua, LI Yun-feng
Journal of Integrative Agriculture    2019, 18 (12): 2673-2681.   DOI: 10.1016/S2095-3119(19)62652-3
摘要164)      PDF    收藏
Rice (Oryza sativa L.) is one of the most important food crops worldwide and a model monocot plant for gene function analysis, so it is an ideal system for studying flower development.  This study reports a mutant, named multi-floret spikelet 3 (mfs3), which is related to the spikelet development in rice and derived from the ethylmethane sulfonate (EMS)-treated rice cultivar XIDA 1B.  In mfs3, the main body of palea (bop) was degenerated severely and only glume-like marginal regions of palea (mrp) remained, while other floral organs developed normally, indicating that the palea identity was seriously influenced by the mutation.  It was also observed that the number of floral organs was increased in some spikelets, including 2 lemmas, 4 mrp, 4 lodicules, 8–10 stamens, and 2 pistils, which meant that the spikelet determinacy was lost to some degree in mfs3.  Furthermore, genetic analysis demonstrated that the mfs3 trait was controlled by a single recessive gene.  Using 426 F2 mutants derived from the cross between sterile line 56S and mfs3, the MULTI-FLORET SPIKELET 3 (MFS3) gene was mapped between the molecular markers RM19347 and RM19352 on Chr.6, with a physical distance of 106.3 kb.  Sequencing of candidate genes revealed that an 83-bp fragment loss and a base substitution occurred in the LOC_Os06g04540 gene in the mutant, confirming preliminarily that the LOC_Os06g04540 gene was the MFS3 candidate gene.  Subsequent qPCR analysis showed that the mutation caused the down-regulation of OsMADS1 and FON1 genes, and the up-regulation of OsIDS1 and SNB genes, which are all involved in the regulation of spikelet development.  The MFS3 mutation also significantly reduced the transcription of the REP gene, which is involved in palea development.  These results indicated that the MFS3 gene might be involved in the spikelet meristem determinacy and palea identity by regulating the expression of these related genes.
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7. Phenotypic characterization and fine mapping of mps1, a premature leaf senescence mutant in rice (Oryza sativa L.)
LIU Zhong-xian, CUI Yu, WANG Zhong-wei, XIE Yuan-hua, SANG Xian-chun, YANG Zheng-lin, ZHANG Chang-wei, ZHAO Fang-ming, HE Guang-hua, LING Ying-hua
Journal of Integrative Agriculture    2016, 15 (9): 1944-1954.   DOI: 10.1016/S2095-3119(15)61279-5
摘要1857)      PDF    收藏
   Leaves play a key role in photosynthesis in rice plants. The premature senescence of such plants directly reduces the accumulation of photosynthetic products and also affects yield and grain quality significantly and negatively. A novel premature senescence mutant, mps1 (mid-late stage premature senescence 1), was identified from a mutant library consisting of ethyl methane sulfonate (EMS) induced descendants of Jinhui 10, an elite indica restorer line of rice. The mutant allele, mps1, caused no phenotypic differences from the wild type (WT), Jinhui 10, but drove the leaves to turn yellow when mutant plants grew to the tillering stage, and accelerated leaf senescence from the filling stage to final maturation. We characterized the agronomic traits, content of photosynthetic pigments and photosynthetic efficiency of mps1 and WT, and fine-mapped MPS1. The results showed that the MPS1-drove premature phenotype appeared initially on the leaf tips at the late tillering stage and extended to the middle of leaves during the maturing stage. Compared to the WT, significant differences were observed among traits of the number of grains per panicle (–31.7%) and effective number of grains per panicle (–38.5%) of mps1 individuals. Chlorophyll contents among the first leaf from the top (Top 1st), the second leaf from the top (Top 2nd) and the third leaf from the top (Top 3rd) of mps1 were significantly lower than those of WT (P<0.05), and the levels of photosynthetic efficiency from Top 1st to the forth leaf from the top (Top 4th) of mps1 were significantly lower than those of WT (P<0.01). Results from the genetic analysis indicated that the premature senescence of mps1 is controlled by a recessive nuclear gene, and this locus, MPS1 is located in a 37.4-kb physical interval between the markers Indel145 and Indel149 on chromosome 6. Genomic annotation suggested eight open reading frames (ORFs) within this physical region. All of these results will provide informative references for the further researches involving functional analyses and molecular mechanism exploring of MPS1 in rice.
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8. YGL9, encoding the putative chloroplast signal recognition particle 43 kDa protein in rice, is involved in chloroplast development
WANG Zhong-wei, ZHANG Tian-quan, XING Ya-di, ZENG Xiao-qin, WANG Ling, LIU Zhong-xian, SHI Jun-qiong, ZHU Xiao-yan, MA Ling, LI Yun-feng, LING Ying-hua, SANG Xian-chun, HE Guang-hua
Journal of Integrative Agriculture    2016, 15 (05): 944-953.   DOI: 10.1016/S2095-3119(15)61310-7
摘要1656)      PDF    收藏
    The nuclear-encoded light-harvesting chlorophyll a/b-binding proteins (LHCPs) are specifically translocated from the stroma into the thylakoid membrane through the chloroplast signal recognition particle (cpSRP) pathway. The cpSRP is composed of a cpSRP43 protein and a cpSRP54 protein, and it forms a soluble transit complex with LHCP in the chloroplast stroma. Here, we identified the YGL9 gene that is predicted to encode the probable rice cpSRP43 protein from a rice yellow-green leaf mutant. A phylogenetic tree showed that an important conserved protein family, cpSRP43, is present in almost all green photosynthetic organisms such as higher plants and green algae. Sequence analysis showed that YGL9 comprises a chloroplast transit peptide, three chromodomains and four ankyrin repeats, and the chromodomains and ankyrin repeats are probably involved in protein-protein interactions. Subcellular localization showed that YGL9 is localized in the chloroplast. Expression pattern analysis indicated that YGL9 is mainly expressed in green leaf sheaths and leaves. Quantitative real-time PCR analysis showed that the expression levels of genes associated with pigment metabolism, chloroplast development and photosynthesis were distinctly affected in the ygl9 mutant. These results indicated that YGL9 is possibly involved in pigment metabolism, chloroplast development and photosynthesis in rice.
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9. Identification and Gene Mapping of a multi-floret spikelet 1 (mfs1) Mutant Associated with Spikelet Development in Rice
REN De-yong*, LI Yun-feng*, WANG Zeng, XU Fang-fang, GUO Shuang, ZHAO Fang-ming, SANG Xianchun, LING ing-hua, HE Guang-hua
Journal of Integrative Agriculture    2012, 12 (10): 1574-1579.   DOI: 10.1016/S1671-2927(00)8690
摘要1647)      PDF    收藏
In this study, a rice spikelet mutant, multi-floret spikelet 1 (mfs1), which was derived from ethylmethane sulfonate (EMS)- treated Jinhui 10 (Oryza sativa L. ssp. indica) exhibited pleiotropic defects in spikelet development. The mfs1 spikelet displayed degenerated the empty glume, elongated the rachilla, the extra lemma-like organ and degraded the palea. Additionally, mfs1 flowers produced varied numbers of inner floral organs. The genetic analysis revealed that the mutational trait was controlled by a single recessive gene. With 401 recessive individuals from the F2 segregation population, the MFS1 gene was finally mapped on chromosome 5, an approximate 350 kb region. The present study will be useful for cloning and functional analysis of MFS1, which would facilitate understanding of the molecular mechanism involved in spikelet development in rice.
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