王学德
植物形态发生与发育的分子遗传学和育种学,侧重于棉花纤维细胞和雄配子细胞发生与发育的机理研究,及其在棉花育种中的应用研究
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- 姓名:王学德
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学术头衔:
博士生导师
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学科领域:
海洋化学
- 研究兴趣:植物形态发生与发育的分子遗传学和育种学,侧重于棉花纤维细胞和雄配子细胞发生与发育的机理研究,及其在棉花育种中的应用研究
王学德教授,男,博士,教授,博士生导师,“浙江省跨世纪学术和技术带头人”。1982年毕业于原浙江农业大学作物遗传育种专业,获学士学位;1987年和1994年在南京农业大学作物遗传育种专业分别获硕士和博士学位,1995-1996年在武汉大学遗传学专业从事分子生物学方面的博士后科研工作,1997年起在浙江大学任教,2003-2004年美国加州大学戴维斯分校高级访问学者。研究领域:植物形态发生与发育的分子遗传学和育种学,侧重于棉花纤维细胞和雄配子细胞发生与发育的机理研究,及其在棉花育种中的应用研究。近年来,承担国家和省部级以上的科学研究项目10项,发表过与此相关的学术论文47篇,获省(部)级以上的研究成果奖7项,申报发明专利5项。研究方向主要有两个,其一是棉花细胞质雄性不育的研究,其二是棉花纤维品质形成的机理研究。棉花细胞质雄性不育的研究已历时15余年(1988-),在遗传学、细胞学、分子生物学和育种学等方面进行过较系统的研究,近年取得突破性进展。用转基因技术将“谷胱甘肽S转移酶(Glutathione S-transferase, GST)基因”导入到待改良的恢复系中,育成了一个转基因强恢复系——“浙大强恢”,克服了30年以来一直难以克服的恢复系对不育系恢复力不强的缺陷,组配出一批三系杂交棉组合,如“浙杂166”、“棕杂1号”和“中标1号”等,在生产上已较大面积应用。“转基因三系杂交棉育种的方法”和“提高三系杂交棉制种产量的方法”两项技术已被授予中国发明专利。具有我国知识产权的转基因三系杂交棉,为杂交棉种子产业化的实施,创造了条件。
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王学德, 李悦有
,-0001,():
-1年11月30日
对白色棉、棕色棉和绿色棉纤维进行了电镜观察,结果表明:三种颜色纤维的结构由外到内依次为初生壁、次生壁和中腔.在白色纤维的超微结构中未发现染色较深的物质;在绿色棉纤维中,纤维的次生壁内层存在大量染色深的条纹,这些条纹可能是色素沉积的结果,类似沉积的日轮;在棕色棉纤维的中腔内,存在大量染色深的物质,而且这些物质主要沉积在中腔壁上,这些染色较深的物质可能是棕色棉纤维的色素物质,同时发现棕色棉纤维具有分叉结构。与白色纤维比较,彩色纤维表现出的品质较差,可能与纤维素含量低及色素物质的沉积有关。
彩色棉, 纤维, 超微结构
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【期刊论文】Cloning of fiber-specific cDNAs and their structural variations in four fiber mutants*
王学德, Wang Xuede** Zhu Yuxian① Ji Daofan Jiang Shuli Li Yueyou
,-0001,():
-1年11月30日
A mRNA preferentially expressed in cotton fiber was cloned from fiber total RNA of normal upland cotton TM-1 (wild-type) by using RT-PCR and corresponding cDNA (signed as TM-E6) was sequenced. TM-E6 gene had no intron and contained an open reading frame 771 bp long, and might encode a peptide of 246 amino acids. Other four genes, Fl-E6, Li-E6, N-E6 and Bl-E6, which were homologous to TM-E6 gene, also were isolated from four fiber mutants of Fiberless Xu-Zhou 142, Ligon lintless, Naked seed and Brown lint, respectively. Sequence analysis of each of these mutant genes revealed many variations in structure and nucleotide composition of gene when compared with sequence of TM-E6 gene. (1). There was a changeable repetitive segment in which GGCTCA (Gly-Ser) was repeated from three to five times between the 82nd and the 93rd codon in different mutant genes. Since the change of Gly-Ser repetitive segment occurred not only in the mutants but also in the wild-type cotton, the repeat frequency might be not associated with the mutation of fiber characteristics. (2). Among these four mutant genes, the percentage of changed codons was 7.05% in Fl-E6, 4.98% in Li-E6, and 4.15% in N-E6 and Bl-E6. It seems that the percentage of changed codons in E6 sequence was positively correlated to the degree of fiber morphological variation. (3). E6 polypeptides of Two long-fiberless mutants (Fiberless Xu-Zhou 142 and ligon lintless) contained high similar (99.4%) variation in the region of ~174 amino acids from N-terminus, and those of shor-fiberlees mutants (Fiberless Xu-Zhou and naked seed) revealed identical variation in the region from 116th to 220th amino acid. It also seems that there was a parallel relation between E6 protein variation and fiber phenotype mutation. (4). Li-E6 and Bl-E6 gene also expressed at low level in seed coat besides at high level in fiber.
cotton fiber mutant fiber mRAN
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【期刊论文】Breeding for high yield and fiber quality in colored cotton (Gossypium hirsutum L.)
王学德, X. D. Wang, Y. Dutt, Y. G. Zhu and Y. Y. Li
,-0001,():
-1年11月30日
In the present study, three types of color fiber cottons i.e. white, brown and green were compared for their fiber quality and yield. The comparison of fiber quality suggested that color fiber cotton was inferior as compared to white fiber cotton. To know the effect of cellulose, mineral elements (N, P and K) and pH of fiber cells on quality of fiber, these components were studied at different fiber cell development stages in all the three fiber cotton types. The cellulose content is closely associated with quality of fiber. The higher fiber quality of white fiber cotton might be due to the high cellulose content in it as compared to colored fiber cotton. A rapid and slow decrease in pH in white and colored cottons, respectively, might bring some effects in fiber elongation. Among the mineral contents, Potash (K) element positively correlated with the fiber quality traits. The pigment development in brown and green fiber cottons was not similar. In green fiber cotton it take more time to become deep as compared to brown fiber cotton. Ultimately, the possible strategy for improvement in quality of color fiber cotton was discussed. The results of heterosis in colored fiber cotton suggested that heterosis approach could improve yield and quality of colored fiber cotton. In the present study, the hybrid between ZJU12A
color cotton breeding,, Gossypium hirsutum,, fiber cell development,, cellulose,, heterosis,, mineral composition
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【期刊论文】Development of Transgenic Restorer of Cytoplasmic Male Sterility in Upland Cotton
王学德, WANG Xuede, LI Yueyou
,-0001,():
-1年11月30日
A glutathione S-transferase (GST) gene has been introduced into restorers of cytoplasmic male sterility in upland cotton (Gossypium hirsutum L.) using Agrobacterium-mediated transformation. A transgenic restorer, signed as 'Zheda strong restorer', which has strong restorability to male sterility, was selected from progeny plants of transformants. When compared with an American restorer 'DES-HAF277', the fertility restorability of 'Zheda strong restorer' to male sterility was been enhanced by 25.8% in the percentage of viable pollens of hybrid (sterile line
Upland cotton, Cytoplasmic male sterility, Transgenic restorer
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王学德, 赵向前, , 王学德*
,-0001,():
-1年11月30日
植物非光合色素主要由类黄酮和类胡萝卜素组成,但彩色棉纤维中色素种类及其结构尚不清楚。经测定,棕色棉(X008)、绿色棉(S029)和白色棉(徐州142)成熟纤维中总黄酮含量分别为4.424mg/g、3.846mg/g和0.375mg/g,没有检测到类胡萝卜素。通过盐酸-镁粉反应、四氢硼钠还原反应、碱反应、醋酸铅反应、氯化铁反应和紫外光谱吸收的分析表明,甲醇室温提取的棕色棉纤维色素可能是黄酮类化合物,B环上带有邻二酚羟基;甲醇室温提取的绿色棉纤维色素除黄酮类化合物外,还含有3-位羟基被糖甙化的黄酮醇类化合物,A、B环上都带有邻二酚羟基。绿色棉纤维色素中还可能含有分子结构类似于芦丁的黄酮醇化合物。
彩色棉, 纤维色素, 类黄酮
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王学德, 赵向前
,-0001,():
-1年11月30日
以棉花细胞质雄性不育系、保持系、恢复系及杂种F1的花瓣、花药、柱头、蜜腺、苞叶和叶片为材料,测定和比较了各材料的总糖、蔗糖和果糖含量,观察和分析了三系杂交棉制种田中蜜蜂造访的数量与制种产量性状的相关性。试验结果表明:不育系的花瓣、苞叶、蜜腺可溶性总糖和蔗糖含量比可育的恢复系、保持系和F1的含量高,并达到显著或极显著水平,而果糖含量差异不明显。其中,不育系花器蔗糖含量是影响蜜蜂造访制种田蜂群量和不育系结铃率的关键因素;选育花器含糖量高的不育系,有利于提高制种田中的蜜蜂数量,增加不育系的结铃率,以及降低其不孕子率,提高制种产量。
不育系, 制种产量, 糖含量, 棉花
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王学德, X. Li
,-0001,():
-1年11月30日
A novel method for the genetic transformation of cotton pollen by means of vacuum infiltration and Agrobacterium-mediated transformation is reported. The acsA and genes, which are involved in cellulose synthesis in Acetobacter xylinum, were transferred into pollen grains of brown cotton with the aim of improving its fiber quality by incorporating useful prokaryotic features into the colored cotton plants. Transformation was carried out in cotton pollen-germinating medium, and transformation was mediated by vector pCAMBIA1301, which contains a reporter gene β-glucuronidase (GUS), a selectable marker gene, hpt, for hygromycin resistance and the genes of interest, acsA and acsB. The integration and expression of acsA, acsB and GUS in the genome of transgenic plants were analyzed with Southern blot hybridization, PCR, histochemical GUS assay and Northern blot hybridization. We found that following pollination on the cotton stigma transformed pollen retained its capability of double-fertilization and that normal cotton seeds were produced in the cotton ovary. Of 1,039 seeds from 312 bolls pollinated with transformed pollen grains, 17 were able to germinate and grow into seedlings for more than 3 weeks in a nutrient medium containing 50mg/l hygromycin; eight of these were transgenic plants integrated with acsA and acsB, yielding a 0.77% transformation rate. Fiber strength and length from the most positive transformants was 15% greater than those of the control (non-transformed), a significant difference, as was cellulose content between the transformed and control plants. Our study suggests that transformation through vacuum infiltration and Agrobacterium mediated transformation can be an efficient way to introduce foreign genes into the cotton pollen grain and that cotton fiber quality can be improved with the incorporation of the prokaryotic genes acsA and acsB.
Cotton
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王学德, CHENG Chaohua, WANG Xuede & NI Xiyuan
Chinese Science Bulletin Vol. 50 No.1 January 2005,-0001,():
-1年11月30日
Lintless mutant is a super-short fiber mutant in upland cotton only 4-8mm in fiber length and also named Ligon cotton controlled by one dominant gene Li1. Fiber ultrastructure of the mutant (Li1) and its wild type (li1) in situ and in vitro was observed under an electron microscope to understand its cytological characteristics during the fiber cell elongation. The results showed that the mutant fiber in situ had thinner cytoplasm, more small vacuoles, less mitochondria, Golgi apparatus and endoplasmic reticula, and there were more starch granules which were free or packed in the amyloplast beside the cell wall than that of wild type. It was indicated that scarcity of functional organelles and disability of transformation from starch to sugar might be associated with the fact that the mutant fiber cell was aborted too early to elongate into normal length. Mutant ovule in some media containing GA3 could produce a kind of huge callus that grew faster than normal ovules. The callus was covered with many white, loose, and semitransparent fiber-like cells that apt to get off from ovule. These fiber-like cells were multicellular fibers generated by cell division and had black dots just like pigment glands in the stem and leaf of cotton. There were lots of micro-tubes beside cytoplasm membrane of the multicellular fiber, which were thought to be primary preparation for second wall deposition of multicellular fiber. It was indicated that GA3 might induce the expression of gene(s) that kept inactive in the field condition and then stimulate the original fiber cell in vitro to undergo division again.
mutant,, ultrastructure,, multicellular fiber.,
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【期刊论文】用IAA和GA3离体诱导棉花短纤维突变体胚珠的纤维伸长1
王学德, 程超华, 王学德*
,-0001,():
-1年11月30日
以棉花等基因系超短纤维突变体(Ligon Li1)及其野生型(Ligon li1)为材料,用胚珠离体培养方法,研究IAA(生长素)和GA3(赤霉酸)与纤维细胞伸长的关系。研究表明:(1)在含单激素IAA或GA3培养基内,离体诱导突变体胚珠产生的纤维长度分别约为1.86mm和2.1mm,比在对照(不含激素)培养基内的纤维长度分别显著增长86%和110%,表明外源激素IAA或GA3对突变体纤维伸长能力的缺损有补偿作用。(2)在同时含有IAA和GA3两种激素的培养基中,突变体和野生型胚珠产生的纤维长度分别为3.41mm和7.23mm,是它们在田间自然条件下纤维长度的76%和24.3%,均显著高于只含一种激素培养基内的纤维长度,这表明IAA与GA3间在诱导纤维伸长上有协同作用,而且,这种作用对突变体的作用大于野生型高达3.1倍,即:在一定程度上能解除突变体纤维伸长受到的阻碍。(3)突变体胚珠在含GA3和IAA培养基内经培养21d后,胚珠IAA和GA3平均含量,虽稍高于对照培养基内野生型胚珠的含量,但明显小于GA3和IAA培养基内野生型胚珠的平均含量,而且其纤维长度也显著小于野生型的纤维长度。说明在离体培养条件下,突变体的纤维伸长比野生型需更高的胚珠GA3和IAA水平,但由于突变体在遗传上的缺损,外源GA3和IAA尚不能完全解除突变体胚珠纤维伸长受到的限制。这可能与突变体胚珠利用外源激素能力不及野生型胚珠有关。
IAA (, 生长素), , GA3 (, 赤霉酸), , 纤维伸长, 胚珠培养
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【期刊论文】A suppressed gene in integument cells of a fiberless seed mutant in upland cotton
王学德, Wang Xuede * Jiang Shuli Li Yueyou Xu Yanglong
,-0001,():
-1年11月30日
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.
upland cotton
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