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A suppressed gene in integument cells of a fiberless seed mutant in upland cotton

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Journal of Zhejiang University Science 2002年第5期3卷 594-599页

作者：王学德蒋淑丽等 collegeofagricultureandbiotechnology ZhejiangUniversityZhejiangHangzhou310029China

A fiberless seed mutant (fl) was identified in a commercial cotton ( Gossypium hirsutum L.) variety Xu Zhou 142 (FL). This phenotype is associated with lack of fiber cell initiation in the outer integument of the ovule, as was characterized by analysis of genes related to fiber differentiation and development. Two genes, fl E6 and FL E6, were cloned from fl integument cells and FL fiber or integument cells, respectively. Compared with FL E6, fl E6 showed a dramatic change in nucleotide sequence: (1) FL E6 contained a tandem repetitive sequence in which GGCTCA (Gly Ser) is repeated five times between the 82nd and the 93rd codon from the first ATG codon, while in fl E6 the same sequence is repeated four times; (2) The fl E6 gene encodes a polypeptide of 241 amino acids but lacks two codons between the 90th and 93rd codon and three between the 171st and 174th relative to FL E6; (3) There are also 12 nucleotide substitutions which would result in 7 amino acid differences between fl E6 and FL E6. Analysis of RT PCR and Northern Blot showed that expression of the fl E6 gene is suppressed in the fl integument cells, but highly expressed in FL fiber cells. The difference between fl E6 and FL E6 may be associated with lower expression of fl E6 in the fl integument cells. Searches of protein databases with the FL E6 gene sequence showed similarity to the protein backbones of two arabinogalactan proteins (AGPs), one from the filtrate of suspension cultured cells of Pyrus communis (AGPPc2) and the other from Nicotiana alata (AGPNa2). Although the function of the FL E6 protein in differentiation and development of cotton fiber cells is not known, the data indicate that the mutation of fl E6 gene from FL E6 gene may inhibit the fiber cell initiation from epidermal cells of the outer integument of the ovule.

关键词： Upland cotton Fiberless seed mutant Integument cells cDNA Arabinogalactan proteins

Inheritance of the Pale—Green Leaf Marker and Sterility Trait in Photo—Thermo Sensitive Genic Male Sterile Rice

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Agricultural Sciences in China 2003年第3期2卷 245-249页

作者： TIANZhen－tao XUEQing－zhong DepartmentofAgronomy CollegeofAgricultureandBiotechnologyZhejiangUniversityHangzhou310029P.R.China DepartmentofAgronomy CollegeofAgricultureandBiotechnologyZhejiangUniversityHangzhou310029P.R.Ch

P/TGMS (photo-thermo sensitive genie male sterility) lines with pale-green leaf color have been developed in japonica rice. The marker trait is used as an assistant selection in the production of the two-lines system hybrid rice for the improvement of F, seed purity. A joint inheritance study of both leaf color and male sterility is presented for P/TGMS line with pale-green leaf color. The segregation ratios for leaf color in the F2 populations of the three crosses showed 13 : 3 and 15 : 1 at early and late sowing stages (April 26 and June 23) respectively, implying that the leaf color is controlled by two genes with fertility gene as dominant. Sterility level is higher in the early sowing stage than that in the late sowing. The inducement of male sterility is closely related to longer day-length and higher temperature at the developmental stages of young panicle. The genes to govern the leaf color and male fertility are inherited independently.

关键词： Rice Pale-green leaf Male sterility Agronomic trait P/TGMS

Detection and Analysis of High Temperature Sensitivity of TGMS Lines in Rice Using AMMI Model

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Agricultural Sciences in China 2004年第9期3卷 671-677页

作者： FULi-zhong XUEQing-zhong DepartmentofAgronomy CollegeofAgricultureandBiotechnologyZhejiangUniversityHangzhou310029P.R.China

With the AMMI (additive main effects and multiplicative interaction) analysis model, thedetermination of the sensitivity to temperature among different TGMS (thermo-sensitivegenic male sterile) lines was performed. To assess the genetic differences due to hightemperature stress at the fertility-sensitive stage (10-20d before heading), sevengenotypes (six TGMS lines and the control Pei-Ai64S) were grown from May 4 at sevendifferent stages with 10d intervals. The temperatures at the fertility-sensitive stagesinvolved twelve levels from℃ under the regime natural conditions in Hangzhou,China. There was considerable variation in pollen fertility among genotypes in responseto high temperature. Five genotypes identified as TGMS lines as their percentages offertile pollens were lower than or close to that of the control except for the unstableline RTS19 (V6). When the temperatures at the fertility-sensitive stage were at Ⅰ-Ⅳ,Ⅴ-Ⅵ and Ⅶ-Ⅻ, the percentages of fertile pollens varied in the ranges of 46.46-48.49%,19.62-22.79% and 3.49-5.87%, respectively. The critical temperatures of sterility andfertility in the five TGMS lines were 25.1 and 23.0℃, respectively. Considering theamounts and directions of main effect and their IPCA (interaction principal componentsanalysis), we can classify the lines and temperature levels into different groups, anddescribe the characteristics of genotypetemperature interaction, offering the informationand tools for the development and utility of thermo-sensitive male sterile *** TGMS rice lines with their reproductive sensitivity to high temperature that canbe screened using the AMMI model may add valuable germplasm to the breeding program ofhybrid rice.

关键词： Rice AMMI (additive main effects and multiplicative interaction) TGMS (thermo- sensitive genic male sterile) Fertility

Cytoplasmic male sterility of tuber mustard is associated with the alternative spliced mitochondrial T gene transcripts

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Chinese Science Bulletin 2004年第23期49卷 2481-2486页

作者： PEIYanxi CHENZhujun CAOJiashu CHENXueiun LIUXiaohui CollegeofLifeScienceandTechnology ShanxiUniversityTaiyuan030006China collegeofagricultureandbiotechnology ZhejiangUniversityHangzhou310029China KeyBiotechnologyLabofYunnanTobaccoScienceAcademy Kunming650106China

Two transcripts of T gene, T1170 and T1243, were obtained from the mitochondrial cDNA of tuber mus- tard CMS line. T1243 was a transcript with an intron unspliced, which has the basic characteristics of type Ⅱ intron. The expressions of the two transcripts were analyzed by reverse transcription PCR (RT-PCR). The results showed that, at seedling stage, the expression of T gene was mainly in the form of T1170 but decreased with the development gradually, while the expression abundance of another tran- script, T1243, increased gradually. The T1243 was prevalent at the profuse flowering stage. The expression pattern was confirmed by Northern blot analysis. These results suggested that the alternative spliced mitochondrial T gene transcripts were related to CMS of tuber mustard.

关键词：雄性不育基因细胞质芥末块茎基因 DNA

Interspecific Hybridization of Brassica campestris × *** Through Ovary Culture

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Agricultural Sciences in China 2004年第2期3卷 124-128页

作者： ZHANGGuo-qing SONGWen-jian TANGGui-xiang ZHOUWei-jun DepartmentofAgronomy CollegeofAgricultureandBiotechnologyZhejiangUniversityHangzhou310029P.R.China

Using three varieties of Brassica campestris, Hauarad (708), Maoshan-3 (714) and Youbai (715),as the maternal plants and one variety of Brassica oleracea Jingfeng-1 (6012) as paternalplants, crosses were made to produce interspecific hybrids through ovary culture *** ovaries from the cross between B. *** (7086012 and 7146012) werecultured and ovary culture was more effective in terms of obtained seeds when ovaries werecultured in vitro at 9 d after pollination (DAP). While for the cross of 7156012, it wasbetter when ovaries in vitro cultured at 12 DAP. Among three cross combinations, the cross of7146012 showed the best response and 43 seeds per ovary were obtained. Among the mediastudied, the ovaries from the cross of 7086012 cultured on MS media supplemented with 3.0 mgL-1 BA0.1 mg L-1 NAA showed better response, and its rate of seeds per ovary reached 44.0%.While the ovaries from the other two crosses (7146012 and 7156012) showed the best responsewhen cultured on B5 media supplemented with 3.0 mg L-1 BA + 0.2 mg L-1 NAA, and the rates of seedsper ovary reached 72.0 and 60.0%, respectively. All seeds obtained from the three crosscombinations were cultured on the MS media supplemented with 1.0 mg L-1 BA + 0.05 mg L-1 NAA,and the seeds from the cross of 7156012 showed the best germination response and thepercentage of germinations reached 66.7%. The regenerated plantlets were obtained from theseseedlings after cultured on the MS media supplemented with 0.05 mg L-1 NAA. Cytological studyshowed that these regenerated plants were all true hybrids of ***.

关键词： Brassica campestris Brassica oleracea Ovary culture Plant regeneration Cytological identification

Enhancing rice resistance to fungal pathogens by transformation with cell wall degrading enzyme genes from Trichoderma atroviride

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Journal of Zhejiang University Science 2004年第2期5卷 133-136页

作者：刘梅孙宗修朱洁徐同 HARMANGaryE LORITOMatteo collegeofagricultureandbiotechnology ZhejiangUniversityHangzhou310029China KeyLaboratoryforRiceBiology MinistryofAgricultureChinaNationalRiceResearchInstituteHangzhou310006China DepartmentofHorticulturalSciences CornellUniversityGenevaUSA InstituteofPlantPathology NapoliUniversityTriticiItaly

Three genes encoding for fungal cell wall degrading enzymes (CWDEs), ech42, nag70 and gluc78 from the biocontrol fungus Trichoderma atroviride were inserted into the binary vector pCAMBIA1305.2 singly and in all possible combinations and transformed to rice plants. More than 1800 independently regenerated plantlets in seven different populations (for each of the three genes and each of the four gene combinations) were obtained. The ech42 gene encoding for an endochitinase increased resistance to sheath blight caused by Rhizoctonia solani, while the exochitinase-encoding gene, nag70, had lesser effect. The expression level of endochitinase but exochitinase was correlated with disease resistance. Nevertheless, exochitinase enhanced the effect of endochitinase on disease resistance when the two genes co-expressed in transgenics. Resistance to Magnaporthe grisea was found in all kinds of regenerated plants including that with single gluc78. A few lines expressing either ech42 or nag70 gene were immune to the disease. Transgenic plants are being tested to further evaluate disease resistance at field level. This is the first report of multiple of expression of genes encoding CWDEs from Trichoderma atroviride that result in resistance to blast and sheath blight in rice.

关键词： Agrobacterium-mediated transformation Rice sheath blight Rice blast Trichoderma atroviride Chitinase Glucanase

Variation in Gelatinization Characteristics of Storage Root Starch During Sweetpotato Growth and Development

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Agricultural Sciences in China 2005年第6期4卷 436-442页

作者： HUANGHua-hong LUGuo-quan ZHENGYi-fan collegeofagricultureandbiotechnology ZhejiangUniversityHangzhou310029P.R.China ZhejiangUniversityofTechnology Hangzhou310032P.R.China ZhejiangForestryCollege Lin＇an311300P.R.China／／CollegeofAgricultureandBiotechnologyZhejiangUniversityHangzhou310029P.R.China

The differential scanning calorimetry (DSC) and rapid visco-analyzer (RVA) were used to determine the starch gelatinizationcharacteristics during the growth of three sweetpotato cultivars. The results showed that the starch contents of threesweetpotato cultivars all decreased as growth progressed. Changes of the amylose contents differing in harvesting datesshould be discriminated according to the cultivars. At the early harvests amylose contents of Xushu18 and Zheda9201were relatively high, but those of Zhe3449 were low. As the growth duration of sweetpotatoes prolonged, the peaks ofDSC thermograms tended to occur at a low temperature and not to be so obstrusive with the increased width of the *** decreases were observed, values of onset, peak and conclusion temperatures, as well as enthalpy of phasetransitions, as growth time lengthened. The peak viscosities, as determined through RVA, showed a rising tendency asgrowth progressed. In addition, statistical analysis revealed that there were correlations between the amylose content andgelatinization characteristics to some extent, which were affected by genotypes evidently.

关键词： Sweetpotato Starch gelatinization characteristics Growth duration DSC RVA

Effect of the Flanking Sequence Architecture of Translation Initiation AUG Codon on Gene Expression Level in Rice

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Agricultural Sciences in China 2004年第6期3卷 401-408页

作者： LIUQing-po TANJun XUEQing-zhong DepartmentofAgronomy CollegeofAgricultureandBiotechnologyZhejiangUniversityHangzhou310029P.R.China HangzhouGenomicsInstitute Hangzhou310007P.R.China

The relationship between the codon usage bias, gene expression level and the AUG context(from -20 to +6 positions relative to the initiator AUG codon) was examined in 541unigene sequences of rice. A significant correlation for CAI values (codon adaptationindex) was observed at five nucleotide positions (-19, -18, -9, -4, +5), eight (-19, -18,-14, -9, -6, -4, -1, +5) for CPP (codon preference parameter), and seven (-18, -16, -15,-9, -7, -1, +6) for mRNA abundance in the flanking sequence of the initiator AUG codonrespectively, but a significantly positive correlation for both CAI and CPP at twopositions (-4 and +5), indicating that both those positions are evolutionally under thenatural selection constraint at the translational level. By site-directed mutagenesis atseven specific positions (-18, -16, -15, -9, -7, -1 and +6) for allergenic protein thathad the highest mRNA abundance in this study, its expression level decreased dramatically63.3 and 72.5% respectively, indicating the importance of those 7 positions for geneexpression. A highly positive correlation (r=0.625, P<0.01) between AUGCAI and GCcontent in the flanking sequence of the initiator AUG codon showed a more effectivehigher GC content on translation initiation efficiency. The strong preference for G orC at those 8 positions (-6, -5, -3, -2, -1, +4, +5 and +6) in the AUG context suggestedthat an important factor in modulation of the translation efficiency, as well assynonymous codon usage bias, particularly in highly expressed genes.

关键词： Rice AUG context Codon usage bias Gene expression level Simulation of point mutation

Effect of Grain Position within a Panicle and Variety on As,Cd,Cr,Ni,Pb Concentrations in japonica Rice

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Rice science 2005年第1期12卷 48-56页

作者： CHENGWang-da ZHANGGuo-ping YAOHai-gen TANGMei-ling JiaxingAcademyofAgriculturalSciences Jiaxing314016China DepartmentofAgronomy CollegeofAgricultureandBiotechnologyZhejiangUniversityHangzhou310029China DepartmentofAgronomy CollegeofAgricultureandBiotechnologyZhejiangUniversityHangzhou310029China／／JiaxingAcademyofAgriculturalSciencesJiaxing314016China

Two japonica rice varieties, a compact panicle (CP) Xiushui 63 and a loose panicle (LP) Xiushui 11 were grown in a slightly contaminated paddy field, to determine the effect of grain positions within a panicle and variety on grain As, Cd, Cr, Ni and Pb concentrations. There was a significant difference in grain As, Cd, Cr, Ni and Pb concentrations among grain positions, and the extent of the difference was variety- and metal-specific; Xiushui 63 showed greater difference than Xiushui 11, and As, Cd, Cr and Pb being greater than Ni. The top grains (TG) of a panicle had higher As, Cd and Ni concentrations than the middle ones (MG), and the bottom ones (BG) contained the lowest. With regard to Cr and Pb, the case was opposite. Significantly positive correlations were found between grain weight and As, Cd, Ni concentrations, but there was a negative correlation between grain weight and Cr, and Pb concentrations. There was a remarkable variation in grain filling pattern among grains within a panicle, and between Xiushui 63 and Xiushui 11. In comparison with MG and TG, BG had the lowest grain filling rate and grain weight, leading to obvious difference in grain weight among grains within a panicle, and a greater difference for Xiushui 63 than for Xiushui 11. The regression analysis showed that grain As and Cd concentrations were positively correlated with the maximum grain filling rate (GRm),while negatively correlated with the required time for reaching the maximum grain filling rate (Tpol). Concerning Cr and Pb, the case was just reverse. It is suggested that As and Cd accumulation in grains might be accompanied by the carbohydrate accumulations, while Cr and Pb accumulation displayed a different pattern.

关键词： rice (Oryza sativa) grain position heavy metal grain weight grain filling