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1.
Discovery of MicroRNAs Associated with the S Type Cytoplasmic Male Sterility in Maize
YU Jiang-hua, ZHAO Yan-xin, QIN Ya-ting, YUE Bing, ZHENG Yong-lian , XIAO Hai-lin
Journal of Integrative Agriculture 2013, 12 (
2
): 229-238. DOI:
10.1016/S2095-3119(13)60222-1
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MicroRNAs (miRNAs) are endogenous small RNAs that play important regulatory roles in the growth and development processes of plants and animals. In this study, we examined the expression profiles of pollen miRNAs from a maize S type cytoplasmic male sterile line and its fertility restored line. In total, 100 known miRNAs belonging to 20 families and 81 novel miRNAs belonging to 44 families were identified. Two and seven known miRNAs had significant expression difference between the two lines at the level of P-value<0.01 and 0.011.5 fold expression difference were verified by stem-loop RT-qPCR. Gene Ontology analysis of miRNA target genes revealed that these genes mainly participated in the transcriptional regulation processes.
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2.
Morphological, Biochemical and Genetic Analysis of a Brittle Stalk Mutant of Maize Inserted by Mutator
FU Xue-qian, FENG Jing, YU Bin, GAO You-jun, ZHENG Yong-lian, YUE Bing
Journal of Integrative Agriculture 2013, 12 (
1
): 12-18. DOI:
10.1016/S2095-3119(13)60200-2
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1531
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Mutants on stalk strength are essential materials for the studies on the formation of plant cell wall. In this study, a brittle stalk mutant of maize, designated as Bk-x, was screened from a Mutator inserted mutant library. At the germination and early seedling stage, the mutant plants were indistinguishable from the normal ones. However, all of the plant organs were brittle after the 5th-leaf stage and remained brittle throughout the rest of the growing period. Microstructure observation showed that the cell wall in vascular bundle sheath of Bk-x was thinner than that in normal plants. The leaf mechanical strength in Bk-x was 77.9% of that in normal plants growing at Xishuangbanna (BN), Yunnan province and that was 61.7% in Wuhan (WH), Hubei Province, China. The proportion of cellulose was 12.3% in Bk-x, which was significantly lower than that in normal plants (26.7%), while the soluble sugar content was 36.1% in Bk-x, which is significantly higher than that in normal plants (12.4%). Genetic analysis using two F2 populations and one F2:3 families demonstrated that the trait of brittle stalk is controlled by a single recessive gene.
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3.
Using the Phosphomannose Isomerase (PMI) Gene from Saccharomyces cerevisiae for Selection in Rice Transformation
WANG Tao, LIU Liang-yu, TANG Yong-yan, ZHANG Xiao-bo, ZHANG Mei-dong, ZHENG Yong-lian, ZHANG Fang-dong
Journal of Integrative Agriculture 2012, 12 (
9
): 1391-1398. DOI:
10.1016/S1671-2927(00)8670
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The phosphomannose isomerase (PMI) gene from Saccharomyces cerevisiae acted as selectable marker and mannose acted as selective agent for the production of transgenic plants of rice (Oryza sativa L.) via Agrobacterium-mediated transformation. The concentration of mannose during the selection was stepwise increased, 5 g L-1 mannose combined with 15 g L-1 sucrose and 500 mg L-1 cefotaxime was used in the initial selection stage, then the concentration of mannose was increased to 11 g L-1, the highest transformation rate was 20.0%. The integration of PMI gene was confirmed by PCR, and the result of RT-PCR assay proved that the intron of PMI gene can be excised correctly during RNA splicing. β- Glucuronidase (GUS) activity analysis confirmed the expression of GUS gene. All those means the PMI gene from yeast can be used as a selectable marker in rice transformation.
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4.
Isolating the Mutator Transposable Element Insertional Mutant Gene mio16 ofMaize UsingDoubleSelectedAmplification of Insertion Flanking Fragments (DSAIFF)
ZHONG Wen-juan, ZHANG Mei-dong, YANG Liu-qi, WANG Ming-chun, ZHENG Yong-lian, YANG Wenpeng GAO You-jun
Journal of Integrative Agriculture 2012, 12 (
10
): 1592-1600. DOI:
10.1016/S1671-2927(00)8692
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1452
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Mutator transposable element (Mu) has been used as an effective tool to clone maize (Zea mays L.) genes. One opaque endosperm mutant (mio16) was identified in a pool of Mu inserted mutants. A modified method, termed the double selected amplification of insertion flanking fragments (DSAIFF), was employed to isolate the Mu flanking fragments (MFFs) of mio16. The target site duplications (TSDs) isolated from the Msp I and Mse I digested MFFs had a same 9-bp sequence and were confirmed to be the flanking sequence of one identically inserted gene. Co-segregation analysis suggested that the MFFs were associated with the mutant opaque endosperm, and mio16 was mapped in silico onto the physical position ranged from 229 965021 to 229 965409 bp of the maize chromosome 4.09 bin. The full-length cDNA of the wild-type gene was obtained by an RT-PCR primer-scanning technique, and Mio16 was found to putatively encode a homolog of the Arabidopsis MAP3K delta-1 protein kinase. RT-PCR result the mRNA expression of mio16 region anchored by primers Mu20 and af276 was not interrupted by Mu insertion. Further researches will be done to elucidate how the expression of mio16 is alternated by Mu insertion.
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5.
Conversion of the Statistical Combining Ability into a Genetic Concept
LV Ai-zhi, ZHANG Hao, ZHANG Zu-xin, TAO Yong-sheng, YUE Bing , ZHENG Yong-lian
Journal of Integrative Agriculture 2012, 12 (
1
): 43-52. DOI:
10.1016/S1671-2927(00)8512
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Since the combining ability was proposed in 1942, efforts to uncover the genetic basis underlying this phenomenon have been ongoing for nearly 70 yr, with little success. Some breeding strategies based on evaluation of combining ability have been produced, and are still extensively used in hybrid breeding. In this review, the genetic basis underlying these breeding strategies is discussed, and a potential genetic control of general combining ability (GCA) is postulated. We suggested that GCA and the yields of inbred lines might be genetically controlled by different sets of loci on the maize genome that are transmitted into offspring. Different inbred lines might possess different favorable alleles for GCA. In hybrids, loci involved in multiple pathways, which are directly or indirectly associated with yield performance, might be regulated by GCA loci. In addition, a case of GCA mapping using a set of testcross progeny from introgression lines is provided.
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