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Isolation and Characterization of GmSTY1,a Novel Gene Encoding a Dual-Specificity Protein Kinase in Soybean（ Glycme max L.）

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Journal of Integrative Plant Biology 2006年第7期48卷 857-866页

作者： Zhao-Shi Xu You-Zhi Ma Xian-Guo Cheng Li-Xia Cao Lian-Cheng Li Ming Chen key laboratory of crop genetics and breeding Ministry of Agriculture Institute of Crop Science the Chinese Academy of Agricultural Sciences Beijing 100081 China

Phosphorylation of protein klnases has profound effects on their activity and interaction with other proteins. Tyroslne phosphorylation was reported to be involved in various physiological processes in plants; however, no typical receptor tyrosine kinase has been isolated from plants thus far. Dual-specificity kinases are potentially responsible for the phosphorylation of both tyrosine and serine/threonine of target proteins. A cDNA clone encoding a putative dual-specificity protein kinase was isolated by screening the cDNA GAL4 activation domain （AD） fusion library of soybean （Glycine max L.）, and its entire length was obtained using 5＇-rapid ampUflcatlon of cDNA ends. The predicted polypeptide of 330 amino acid residues, designated as GmSTY1, contains all 11 conserved subdomains, which share common characteristics with both the serine/ threonine and tyroslne protein klnases reported thus far. In addition, three potential N-linked glycosylation sites （NXS/T）, as well as phosphorylation motifs （SXXXS/T）, were observed, suggesting that GmSTY1 may be post-translationally modified. Furthermore, a potential N-myristoylation motif （MGARCSK） was found, suggesting that the GmSTY1 protein could associate with membranes in vivo. Southern blotting analysis revealed a single-copy of GmSTY1 in the genome. Northern blotting analysis showed that this gene was upregulated by drought and salt treatment in a time-dependent manner; however, exogenous abscisic acid （ABA） could not significantly affect the mRNA accumulation of GmSTY1. Interestingly, the transcript of this gene was remarkably downregulated by cold treatment during the early stages of the response, but upregulated later. These results Indicate that the protein kinase was possibly regulated by abiotic stresses in an ABA-independent pathway.

关键词： abiotic stress dual-specificity Glycine max , serine/threonine/tyrosine protein kinase.

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Characterization of Porcine Matrix Metalloproteinase 23 (pMMP-23) Gene and Its Association with Litter Size Traits

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Journal of Integrative Agriculture 2013年第1期12卷 103-109页

作者： NIU Bu-yue LAN Xiao-ming YAN Xiao-hong DI Sheng-wei WANG Yang LI Hai-tao XIONG Yuan- zhu WANG Xi-biao College of Animal Science and Technology Northeast Agricultural University key laboratory of Swine genetics and breeding Ministry of Agriculture/Key Laboratory of Agricultural Animal Genetics Breeding and Reproduction Ministry of Education/Huazhong Agricultural University

The matrix metalloproteinase 23 （MMP-23）, which might play a role in ovulation in mammals, was one of the promising candidate genes for litter size traits in pigs. In the present research, partial sequence of porcine MMP-23 （pMMP-23） gene, including exons 2-8 （GenBank： EU360790）, was obtained. Real-time PCR analysis revealed that pMMP-23 gene was highly expressed in ovary. PCR-Sau3A I-RFLP and PCR-Acc II-RFLP assay were established to detect a C/T mutation in exon 3 （EU360790： g. 269C〉T） and an A/G mutation in exon 4 （EU360790： g. 505A〉G）, respectively. Association study for these two SNPs with litter size was assessed in three independent populations （Minzhu, Landrace I and Landrace II）. Statistical analysis demonstrated that for second and subsequent litters, TT sows produced more TNB than CC pigs in Landrace breed （P〈0.05） at g. 269C〉T locus, and the additive effect was significant （P〈0.05）; GG sows produced more TNB and NBA than AA pigs in Minzhu （P〈0.01） and Landrace breeds （P〈0.05） at g. 505A〉G locus, and the additive effect was significant （P〈0.01 or P〈0.05）. Our study suggested that the pMMP-23 gene might be a novel candidate gene for litter size traits, and g. 505A〉G locus might be a useful molecular marker for marker assisted selection （MAS）.

关键词： pig MMP-23 molecular cloning expression litter size

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Fine-tuning the spatial allocation of phytohormones for smart canopy, smart breeding, and smart agriculture

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Science China Life Sciences 2025年第1期 309-310页

作者： Tao Zhou Lin Li National key laboratory of crop Genetic Improvement Huazhong Agricultural University HuBei HongShan laboratory

The dual challenges of global climate change and population growth have increased demand for higher crop yields per unit area. The innovative technology of smart agriculture aims to synergize breeding, cultivation, and production through the application of biological and informatics technologies. Historically, the “Green Revolution” introduced semidwarf varieties of certain key crops by utilizing mutations affecting gibberellin pathways, which significantly increased planting densities and yields.

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Isolation,Sequence Analysis and Expression Profile of a Novel Swine Gene Differentially Expressed in the Longissimus Dorsi Muscle Tissues from LandracexLarge White Cross-combination

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Acta Biochimica et Biophysica Sinica 2005年第3期37卷 186-191页

作者： Yong-Gang LIU Yuan-Zhu XIONG Chang-Yan DENG key laboratory of Swine genetics and breeding.ministry of Agriculture,Huazhong agricultural University,Wuhan 430070.China key laboratory of Swine genetics and breeding Ministry of Agriculture Huazhong Agricultural UniversityWuhan 430070 China

The mRNA differential display technique was performed to investigate the differences in geneexpression in the Longissimus dorsi muscle tissues from LandracexLarge White *** gene that was differentially expressed was identified using semi-quantitative reverse transcriptase-polymerase chain reaction（RT-PCR）and its complete cDNA sequence was obtained using the rapid amplifi-cation of cDNA ends（RACE）*** nucleotide sequence of the gene is not homologous to any of theknown porcine *** sequence prediction analysis revealed that the open reading frame of this geneencodes a protein of 260 amino acids that contains the putative conserved domain of the carbonic anhydrase,and this protein has high homology with the carbonic anhydrase Ⅲ（CA-Ⅲ）of four species—mouse（91%）,horse（91%）,rat（89%）and human（86%）—so that it can be defined as swine carbonic anhydrase Ⅲ.Thephylogenetic tree analysis revealed that the swine CA-Ⅲ has a closer genetic relationship with the horse CA-Ⅲthan with those of mouse,rat and *** tissue expression analysis indicated that the swine CA-Ⅲ geneis generally expressed in most *** experiment is the first to establish the primary foundation forfurther research on the swine CA-Ⅲ gene.

关键词： mRNA differential display rapid amplification of cDNA ends(RACE) novel gene carbonic anhydrase Ⅲ(CA-Ⅲ)

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Molecular Cloning and Functional Analysis of MRLC2 Differential Expressed in Meishan×Yorkshire F1 Crossbreeds and Their Parents,Meishan Pigs

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Acta Biochimica et Biophysica Sinica 2006年第11期38卷 788-794页

作者： Hong-Tao XIE Ming-Gang LEI Yuan-Zhu XIONG* Chang-Yan DENG Si-Wen JIANG Bo ZUO Feng-E LI De-Quan XU Tao WANG key laboratory of Swine genetics and breeding,ministry of Agriculture,Huazhong agricultural University,Wuhan 430070,China key laboratory of Swine genetics and breeding Ministry of Agriculture Huazhong Agricultural UniversityWuhan 430070 China

In order to detect the molecular basis of heterosis in pigs,suppression subtractivehybridization was carried out to investigate the difference in gene expression in the Longissimus dorsimuscle tissues between Meishan×Yorkshire F1 crossbreeds and their parents,Meishan *** swinemyosin regulatory light chain 2 (MRLC2) gene differentially expressed between the crossbreeds and thepurebreds was isolated and identified using semi-quantitative reverse transcriptase polymerase chain reactionand its complete cDNA sequence was obtained using the rapid amplification of cDNA ends *** sequence of the gene is not homologous to any of the known porcine *** sequenceprediction analysis reveals that the open reading frame of this gene encodes a protein of 172 amino acidscontaining the putative conserved domain of the EF-hand *** predicted amino acid sequenceof porcine MRLC2 protein exhibits 99%,98%,98%,98% and 97% identity with that of cattle,human,dog,rat and mouse,*** homology analysis revealed that the MRLC2 protein was very muchconserved in *** tissue expression analysis indicated that the swine MRLC2 gene is highlyexpressed in muscle,fat,heart,liver,spleen,lung,kidney,stomach,small intestine,ovary and testis,but notexpressed in pancreas.

关键词： MRLC2 gene pig heterosis Longissimus dorsi suppression subtractive hybridization rapid amplification of cDNA end

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Ectopic Overexpression of Wheat Adenosine Diphosphate-ribosylation Factor,TaARF,Increases Growth Rate in Arabidopsis

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Journal of Integrative Plant Biology 2009年第1期51卷 35-44页

作者： Yingyin Yao Zhongfu Ni Jinkun Du Zongfu Han Yan Chen Qingbo Zhang Qixin Sun Department of Plant genetics & breeding and State key laboratory for Agrobiotechnology Key Laboratory of Crop Heterosis and UtilizationMinistry of Education Key Laboratory of Crop Genomics and Genetic Improvement Ministry of Agdculture Beijing Key Laboratory of Crop Genetic Improvement China Agricultural University Beijing 100094 China

Differential gene expression between hybrids and their parents is considered to be associated with heterosis. However, the physiological functions and possible contribution to heterosis of these differentially expressed genes are unknown. We have isolated one hybrid upregulated gene encoding putative wheat ADP-ribosylation factor, designated TaARF. In this study, real-time quantitative reverse transcription-polymerase chain reaction analysis indicated that the TaARF transcript was preferentially expressed in root, node and crown, and the accumulation of TaARF mRNA in hybrid was more than 1.5-fold higher than that in two parents. In order to understand possible roles of the putative wheat ARF gene, TaARF was overexpressed in Arabidopsis, and the transgenic plants were characterized. We show that ectopic overexpression of TaARF in Arabidopsis leads to increased leaf area, increased growth rate and earlier transition to flowering, suggesting that TaARF plays significant roles in growth and development. This study provides evidence demonstrating that TaARF plays important roles in growth and development and we speculate that the upregulated expression of this gene might contribute to the heterosis observed in wheat root and leaf growth.

关键词： adenosine diphosphate-ribosylation factor Arabidopsis ectopic heterosis overexpression wheat.

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Cloning and expression profiles of 15 genes encoding WRKY transcription factor in wheat (Triticum aestivem L.)

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Progress in Natural Science:Materials International 2008年第6期18卷 697-705页

作者： Hualing Wu,Zhongfu Ni,Yingyin Yao,Ganggang Guo,Qixin Sun* Department of Plant genetics&breeding and State key laboratory for Agrobiotechnology,key laboratory of crop Heterosis and Utilization,ministry of Education,key laboratory of crop Genomics and Genetic Improvement,ministry of Agriculture/Beijing key laboratory of crop Genetic Improvement,China agricultural University,Beijing 100094,China Department of Plant genetics & breeding and State key laboratory for Agrobiotechnology Key Laboratory of Crop Heterosis and Utilization Ministry of Education Key Laboratory of Crop Genomics and Genetic Improvement Ministry of Agriculture/Beijing Key Laboratory of Crop Genetic Improvement China Agricultural University Beijing 100094 China

WRKY proteins are involved in various physiological processes,including biotic and abiotic stress responses,hormone responses and ***,no systematic identification,expression and function analysis of WRKY genes in wheat were *** this study,we isolated 15 wheat cDNAs with complete open reading frame(ORF)encoding putative WRKY proteins using in silico *** analysis indicated that the 15 wheat WRKY genes belonged to three major WRKY *** analysis revealed that most genes expressed drastically in leaf,except TaWRKY10 which expressed in crown *** genes were strongly up-reg-ulated with the senescence of *** genes were responsive to low temperature,high temperature,NaCl or PEG ***-over,dierential expression patterns were also observed between wheat hybrid and its parents,and some genes were more responsive to PEG treatment in the *** results demonstrated that wheat WRKY genes are involved in leaf senescing and abiotic *** the changed expression of these WRKY genes in hybrid might contribute to the heterosis by improving the stress tolerance in hybrids.

关键词： Wheat WRKY transcription factor Gene cloning Gene expression Heterosis

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Prediction of VIGS efficiency by the Sfold program and its reliability analysis in Gossypium hirsutum

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Science Bulletin 2016年第7期61卷 543-551页

作者： Xiaoyang Ge Jie Wu Chaojun Zhang Qianhua Wang Yuxia Hou Zuoren Yang Zhaoen Yang Zhenzhen Xu Ye Wang Lili Lu Xueyan Zhang Jinping Hua Fuguang Li State key laboratory of Cotton Biology Institute of Cotton Research Chinese Academy of Agricultural Sciences Department of Plant genetics and breeding Key Laboratory of Crop Heterosis and Utilization of Ministry of Education Beijing Key Laboratory of Crop Genetic Improvement China Agricultural University College of Science China Agricultural University

Genetic transformation in some plant species, including cotton （Gossypium hirsutum）, is hampered by laborious and time-consuming processes and often unachievable. Virus-induced gene silencing （VIGS） by double-stranded RNAs can serve as a reverse-genetics tool to determine gene function. However, knockdown levels vary greatly when using a tobacco rattle virus-based vector that carries different cDNA fragments of a gene. How to choose the optional target fragment for high interference efficiency is very challenging. Addressing this challenge requires increasing the efficacy of small interference RNA （siRNA） in target fragment. Here, we describe a method to assess VIGS efficiency by comparing the following parameters of siRNA in target sequence： the disruptionenergy of the target （△Gdisruption）, the differential stability of siRNA duplex ends （DSSE）, and the internal stability at positions 9-14 of the siRNA antisense strand （AIS）, which are calculated by Sfold program （http：//***. org）. We find that the siRNAs with low mGdisruption, high DSSE and high AIS have high activity and easily result in high VIGS efficiency by experimentally testing the actual knockdown levels of the four target genes, GhPDS, GhCLA1, GhAOS1, and GhCXE1 via choosing different target sequences for each gene. Therefore, the Sfold pro- gram can be used to analyze target sequences when car- rying out VIGS design to increase gene-silencing effects in plants.

关键词： VIGS Sfold program siRNA Targetsequences Gossypium hirsutum

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Mapping QTL affecting the vertical distribution and seed set of soybean [Glycine max(L.) Merr.] pods

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The crop Journal 2019年第5期7卷 694-706页

作者： Shiping Liu Hong Xue Kaixin Zhang Ping Wang Daiqun Su Wenbin Li Shichao Xu Jianan Zhang Zhongying Qi Yanlong Fang Xiyu Li Yue Wang Xiaocui Tian Jie Song Jiajing Wang Chang Yang Sitong Jiang Wen-Xia Li Hailong Ning key laboratory of Soybean Biology Ministry of Education Key Laboratory of Soybean Biology and Breeding/Genetics Ministry of AgricultureNortheast Agricultural University

Number of pods per plant and number of seeds per pod are quantitative,multigenic traits and important components of yield in soybean[Glycine max(L.)Merr.].Pods are distributed unevenly in the upper,middle,and lower segments of the plant and this distribution is affected by sowing date(SD).A population of four-way recombinant inbred lines(FW-RIL),containing 160 F2:8 individuals,was generated from the cross(Kenfeng 14×Kenfeng 15)×(Heinong 48×Kenfeng 19).A linkage map consisting of 275 simple sequence repeat(SSR)markers was used to map quantitative trait loci(QTL)associated with the production of one,two,three,and four seeds per pod in the upper,middle,and lower segments of plants at two SDs,totaling 12 measurements per SD.A wide range of variation in the twelve characteristics was observed among the four parental lines and the FW-RIL population at the two *** effect of SD2(May 17,2016)on pod number was stronger than that of SD1(May 7,2016)because the heritability of each trait in the SD1 experiment was generally greater than that of *** study identified 76 QTL controlling pod number,with the phenotypic variation explained by each QTL ranging from 1.86%to 13.71%.The numbers of QTL controlling one,two,three,and four seeds per pod were 28,23,23,and 23,*** were 30,28,and 28 QTL controlling the pod number in the upper,middle,and lower segments of the plant,***-five QTL were identified at SD1 and 38 QTL were identified at *** QTL were associated with pod-number *** QTL qPNA1–3 was associated with the number of pods containing one seed in the middle segment of the plant at both ***-three QTL were published QTL(common areas existed when integrating on a map GmComposite2003 of Wm82 based on left and right markers).and 13 QTL related to pod number were newly *** results provide a reference for breeders to improve soybean yield by combining advantageous alleles for these *** studies may reveal candi

关键词： Soybean Pod number QTL Sowing date Vertical distribution

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Molecular mapping of QTLs for root response to phosphorus deficiency at seedling stage in wheat (Triticum aestivum L.)

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Progress in Natural Science:Materials International 2007年第10期17卷 1177-1184页

作者： Li Zhenxing, Ni Zhongfu, Peng Huiru, Liu Zhiyong, Nie Xiuling, Xu Shengbao, Liu Gang and Sun Qixin(key laboratory of crop Heterosis and Utilization and State key laboratory for Agrobiotechnology, key laboratory of crop Genomics and Genetic Improvement, Beijing key laboratory of crop Genetic Improvement, Department of Plant genetics & breeding, China agricultural University, Beijing 100094, China) key laboratory of crop Heterosis and Utilization Department of Plant Genetics and Breeding China Agricultural University Beijing 100094 China key laboratory of crop Genomics and Genetic Improvement Department of Plant Genetics and Breeding China Agricultural University Beijing 100094 China Beijing key laboratory of crop Genetic Improvement Department of Plant Genetics and Breeding China Agricultural University Beijing 100094 China

Phosphorus (P) deficiency in the soil is one of the major abiotic stresses that limit plant growth and crop productivity throughout the world. Development of cultivars with improved P-deficiency tolerance is an efficient strategy for sustainable agriculture. Plant roots play an important role in crop growth and development, especially in nutrient uptake and improvement of P-efficiency. Mapping quantitative trait loci (QTLs) for root traits and their response to low P stress at seedling stage will facilitate the development of P-efficient wheat cultivars. In this study, 30 QTLs (LOD>2.0) were mapped for the three root traits, such as root length, root number and root dry matter under different P supply conditions and their response to P-stress. These QTLs were distributed on 14 chromosomes, with each of the 5 QTLs explaining more than 10% phenotype variance. Analyses showed that root traits and their response to P-deficiency were controlled by different QTLs. In addition, alleles with positive effects were separated on both parents, and wheat cultivars with improved P-efficiency could be developed by accumulating these positive effect alleles together.

关键词： QTLs roots wheat seedling RIL P-deficiency.

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