CHANGES IN GROWTH AND GENE EXPRESSION INDUCED BY SULFUR DEFICIENCY IN GARLIC

ABSTRACT

Sulfur deficiency in garlic Allium sativum L. caused a reduction in growth together with chlorosis and necrosis of leaves. Large differences in shoot sulfur and sulphate concentrations between deficient and high sulfur treatments were only observed after 54 days growth. Using the mRNA differential display technique, a novel cDNA was isolated from shoots grown in sulfur depleted nutrient solution for 24 days. This novel cDNA was constitutively expressed in the shoots during further growth in sulfur depleted solution, but it was undetectable following 30 days recovery with sulfur supplementation. The cDNA sequence demonstrated a high degree of identity with a coat protein gene of a garlic latent carlavirus. The results suggest a possible relationship between low plant sulfur status and the induction of a latent carlavirus in garlic.     

兔防御素基因在毕赤酵母中的串连表达及抑菌机理研究

根据毕赤酵母Pichia pastoris密码子偏爱性重编兔防御素基因(Neutrophils peptide-1,pNP-1),设计4条引物搭桥合成.将pNP-1克隆到pPICZα-A载体,使用同尾酶Xho Ⅰ和SalⅠ在重组载体上构建多价串联体[pNP-1(2×)]和[ pNP-1(3×)],分别对这3个基因进行毕...     

兜兰二氢黄烷酮醇-4-还原酶基因的克隆及其表达特性

克隆同色兜兰二氢黄烷酮醇-4-还原酶基因DFR,分析其序列特征,了解其在不同组织部位的表达情况。采用RT-PCR和RACE技术从花中克隆DFR的全长cDNA,用生物信息学方法分析序列特征,并用半定量RT-PCR研究其在不同组织的表达。分离到DFR,该cDNA全长1267 bp,具有1个1074 bp的完整开放阅读框,编码357个氨基酸。序列分析表明,该DFR蛋白含有1个NADB_Rossmann superfamily保守结构域,1个NADP(H)结合域和1个底物特异性结合域,其与文心兰、石斛兰和大花蕙兰等的DFR同源性在75%~80%。系统进化树显示,该DFR蛋白与兰科植物的DFR蛋白亲缘关系比较近。半定量RT-PCR表明,该DFR在成熟花和营养叶中的表达量最高,在子房、苞叶、萼片和花瓣中的表达量相当,在唇瓣中的表达量次之,在蕊柱中的表达量较唇瓣中的低,在花茎中的表达量最低,在根中几乎检测不到。原核表达结果表明,该DFR在大肠杆菌中获得表达。从兜兰中克隆到花色代谢途径中的关键酶基因DFR,该基因可能参与调控花色素的合成。     

稻瘟病菌无毒基因研究进展

水稻与稻瘟菌间存在广泛而特异的相互作用,是研究寄主与病原物互作的重要模式系统。无毒基因是病原物遗传因子,能诱发寄主植物产生抗病性,是稻瘟菌与水稻互作最重要的激发子。为寻求稻瘟病防治的新策略,本研究概述了研究稻瘟病菌无毒基因的意义,归纳了抗性基因与无毒基因互作的3种模式,总结了国内外稻瘟病菌无毒基因的克隆及功能研究,探讨了无毒基因在生产上病害流行预测中的应用,指出利用抗性基因与无毒基因的关系对稻瘟菌分类将会更为科学和实用,应进一步加快稻瘟病菌无毒基因的克隆。     

西瓜抗枯萎病基因同源序列的克隆与分析

本研究根据已克隆的抗枯萎病基因的NBS保守结构域设计了22条上游简并引物和17条下游简并引物,以西瓜抗枯萎病种质PI296341-FR和感枯萎病品种97103为材料,获得了7条来自基因组DNA的RGA序列（GenBank登录号：DQ156558-DQ156564）,均含有NBS保守区的P-环、kinase-2或kinase-3等抗病基因的特征序列结构,所编码的氨基酸序列与已知抗枯萎病基因Fom-2、I2C-1、I2C-2和I2等编码的氨基酸序列表现出11%～72%的同源性,其中来自PI296341-FR的RGA序列175R1与甜瓜抗枯萎病基因Fom-2的同源性最高,为72%。来自PI296341-FR与97103的RGA序列之间同源性较高（73%～97%）,证明了抗病基因在进化上的保守性。     

Factors involved in Agrobacterium tumefaciens-mediated gene transfer into Pinus nigra Arn. ssp. salzmannii (Dunal) Franco

Cotyledons from dissected sterile embryos of salgareño pine (Pinus nigra Arn. ssp. salzmannii (Dunal) Franco) were inoculated with different disarmed Agrobacterium tumefaciens strains harbouring the binary vector p35SGUSint. The transient expression of a β-glucuronidase gene (uidA) was studied, using a histochemical staining procedure. Nineteen days after inoculation, the activity of β-glucuronidase was detected in epidermal and subepidermal layers of cotyledonary explants. The EHA105 strain harbouring a disarmed agropine-type Ti-plasmid (pTiBO542) was the most effective for gene transfer of the uidA gene. The effects of exudates and extracts from 0-day-old embryos on induction of vir gene expression in A. tumefaciens were also examined. The results of this study showed that salgarño pine embryo exudates contain a substance(s) that induce vir gene expression, in similar way to that observed with 100 μM acetosyringone (AS).All these findings suggest that T-DNA processing and transfer might take place when Agrobacterium infects suitable tissues of salgareño pine.     

Suppression subtractive hybridization library construction and identification of epidermal bladder cell related genes in the common ice plant,Mesembryanthemum crystallinum L.

Mesembryanthemum crystallinum L., a halophytic species, displays modified trichomes, epidermal bladder cells (EBC), on the surfaces of its aerial organs. EBCs serve to sequester excessive salt from underlying metabolically active tissues. To elucidate the molecular determinants governing EBC development in the common ice plant, we constructed a cDNA-based suppression subtractive hybridization library and identified genes differentially expressed between the wild-type and the EBC-less mutant. After hybridization, 38 clones were obtained. Among them, 24 clones had homology with plant genes of known functions, whose roles might not be directly related to EBC-morphology, while 14 clones were homologous to genes of unknown functions. After confirmation by northern blot analysis, 12 out of 14 clones of unknown functions were chosen for semi-quantitative RT-PCR analysis, and the results revealed that three clones designated as MW3, MW21, and MW31 preferentially expressed in the EBC-less mutant, whereas the other two designated as WM10 and WM28 preferentially expressed in the wild type. Among these genes, the expression of a putative jasmonate-induced gene, designated as WM28 was completely suppressed in the EBC-mutant. In addition, the deletion of C-box cis-acting element was found in the promoter region of WM28 in the EBC-less mutant. Overexpression of WM28 in Arabidopsis resulted in increased trichome number due to the upregulation of key trichome-related genes GLABRA1 (GL1), and GLABRA3 (GL3). These results demonstrate that WM28 can be an important factor responsible for EBC formation, and also suggest the similarity of developmental mechanism between trichome in Arabidopsis and EBC in common ice plant.     

Effects on flowering and seed yield of dominant alleles at maturity loci E2 and E3 in a Japanese cultivar,Enrei

‘Enrei’ is the second leading variety of soybean (Glycine max (L.) Merr.) in Japan. Its cultivation area is mainly restricted to the Hokuriku region. In order to expand the adaptability of ‘Enrei’, we developed two near-isogenic lines (NILs) of ‘Enrei’ for the dominant alleles controlling late flowering at the maturity loci, E2 and E3, by backcrossing with marker-assisted selection. The resultant NILs and the original variety were evaluated for flowering, maturity, seed productivity and other agronomic traits in five different locations. Expectedly, NILs with E2 or E3 alleles flowered later than the original variety in most locations. These NILs produced comparatively larger plants in all locations. Seed yields were improved by E2 and E3 in the southern location or in late-sowing conditions, whereas the NIL for E2 exhibited almost the same or lower productivity in the northern locations due to higher degrees of lodging. Seed quality-related traits, such as 100-seed weight and protein content, were not significantly different between the original variety and its NILs. These results suggest that the modification of genotypes at maturity loci provides new varieties that are adaptive to environments of different latitudes while retaining almost the same seed quality as that of the original.     

云南花生丛枝植原体secY基因序列分析及结构预测

目的 确定采自元谋地区自然表现丛枝病的小驳骨(Gendarussa vulgaris Nees)植株是否感染植原体。 方法 通过利用植原体16S组和亚组通用或半通用特异性引物分别对植原体16S rRNA、rp和secY基因序列进行PCR扩增、克隆及测序分析;此外还对secY蛋白的蛋白特性及其结构进行了分析和预测。 结果 本研究获得了基因片段长度分别为1 248 bp的16S rRNA基因(nested PCR)、1 171 bp 的rp基因和1 425 bp的secY基因。基于rp和secY基因核苷酸序列的同源性比对及构建的进化树推断的植原体遗传分化关系几乎与16S rRNA基因的推断相一致,均与植原体16S rII-A亚组各株系的遗传进化关系最为接近,但rp和secY基因序列比16S rRNA基因序列能够呈现出更大的遗传变异程度;此外,对secY蛋白进行了生物信息学分析和初步探讨,发现它具有10个明显且分布相对均匀的跨膜螺旋区域,无信号肽。 结论 小驳骨丛枝病(Gendarussa vulgaris witches’-broom phytoplasma,GvWB-YNym)是由植原体侵染而发生,该植原体株系被划分到16S rII-A亚组,相关的候选种为Candidatus Phytoplasma aurantifolia;secY蛋白生物信息参数的分析表明：secY蛋白在感病小驳骨植株中以疏水性稳定跨膜蛋白的形式存在,含10个明显的疏水跨膜区域,该蛋白不存在信号肽。     

新疆杨胰蛋白酶抑制剂基因的克隆与表达

杨树受损伤后能诱导一些基因的表达,其编码的蛋白质可能在杨树的防卫反应中起一定作用.用PCR方法从新疆杨叶片中克隆出一个损伤诱导型Kunitz胰蛋白酶抑制剂基因PaTI1.序列分析表明:此基因不含内含子,其翻译起点上游具有'TATA'和 'CCAAT'等转录控制元件,包含的阅读框架能编码一个长为213个氨基酸的多肽.此多肽与克隆自美洲山杨的PtTI2 和PtTI1氨基酸序列同源性最高,分别为95%和80%,其N端存在一长度为27个氨基酸的信号肽.将此基因以融合蛋白的形式在大肠杆菌中进行表达,纯化后的融合蛋白对胰蛋白酶的活性有抑制作用,每8.5 μg融合蛋白可完全抑制1 μg牛胰蛋白酶的活性.Western blot分析表明融合蛋白与PtTI2特异的抗体之间有明显的血清学反应.