Genetic Transformation of Tobacco with the Trehalose Synthase Gene from Grifola frondosa Fr. Enhances the Resistance to Drought and Salt in Tobacco

Trehalose is a non-reducing disaccharide of glucose that functions as a protectant in the stabilization of biological structures and enhances the tolerance of organisms to abiotic stress. In the present study, we report on the expression of the Grifolafrondosa Fr. trehalose synthase (TSase) gene for manipulating abiotic stress tolerance in tobacco (Nicotiana tabaccum L.). The expression of the transgene was under the control of two tandem copies of the CaMV35S promoter and was transferred into tobacco by Agrobacterium tumefaciens EHA105. Compared with non-transgenic plants, transgenic plants were able to accumulate high levels of products of trehalose, which were increased up to 2.126-2.556 mg/g FW, although levels were undetectable in non-transgenic plants. This level of trehalose in transgenic plants was 400-fold higher than that of transgenic tobacco plants cotransformed with Escherichia coli TPS and TPP on independent expression cassettes, twofold higher than that of transgenic rice plants transformed with a bifunctional fusion gene (TPSP) of the trehalose-6-phosphate (T-6-P) synthase (TPS) and T-6-P phosphatase(TPP) of E. coil, and 12-fold higher than that of transgenic tobacco plants transformed the yeast TPS1 gene.It has been reported that transgenic plants with E. coil TPS and/or TPP were severely stunted and had morphological alterations of their roots. Interestingly, our transgenic plants have obvious morphological changes, including thick and deep-coloured leaves, but show no growth inhibition; moreover, these morphological changes can restore to normal type in T2 progenies. Trehalose accumulation in 35S-35S:TSase plants resulted in increased tolerance to drought and salt, as shown by the results of tests on drought, salt tolerance, and drought physiological indices, such as water content in excised leaves, malondialdehyde content, chlorophyll a and b contents, and the activity of superoxide dismutase and peroxidase in excised leaves. These results suggest that transgenic plants transformed with the TSase gene can accumulate high levels of trehalose and have enhanced tolerance to drought and salt.     

根施甜菜碱对干旱胁迫下小麦幼苗类囊体膜组分和功能的影响

干旱胁迫造成两小麦品系类囊体膜上的单半乳糖脂甘油二酯(MGDG)、双半乳糖脂甘油二酯(DGDG)、磷脂酰胆碱(PG)以及叶绿素含量显著下降,甜菜碱预处理能缓解这些组分的下降.干旱胁迫下,抗旱型小麦品系HF9 70 3的硫代异鼠李糖甘油二酯(SQDG)、反式十六碳-烯酸[16:1(3t)]含量显著上升,MGDG中亚麻酸(18:3)相对含量显著下降;而干旱敏感型品系SN215953则表现为SQDG、16:1(3t)含量显著下降,MGDG中脂肪酸变化不明显,这可能是两个小麦品系抗旱性差异的重要原因之一;甜菜碱处理能显著减小干旱处理与对照之间的差异,且对SN215953的作用较HF9703大.另外,干旱胁迫引起类囊体膜上Ca2 -ATPase活性、Hill反应活性及叶片净光合速率下降,外源甜菜碱能缓解其下降趋势.     

铝胁迫下植物根系的有机酸分泌及其解毒机理

酸性土壤中的铝毒害问题,已成为限制植物生长发育的主要因素之一.耐铝植物通过根系分泌有机酸来解除或减轻铝的毒害是外部解铝毒的重要机制.文章对铝胁迫下植物根系分泌有机酸的种类,有机酸解铝毒机理、解铝毒能力,有机酸分泌方式及调控其分泌的主要因素等相关研究进行综述.     

Presowing Seed Treatment with Cytokinins and Its Effect on Growth, Photosynthetic Rate, Ionic Levels and Yield of Two Wheat Cultivars Differing in Salt Tolerance

The effects of presowing seed treatment with different concentrations of cytokinins （kinetin and benzylaminopurine; 100, 150, and 200 mg/L） on growth, photosynthetic capacity, and ion homeostasis were investigated in two spring wheat （Triticum aestivum L.） cultivars, namely MH-97 （salt sensitive） and Inqlab- 91 （salt tolerant）. Primed and non-primed seeds were sown in a field in which NaC1 salinity of 15 dS/m was developed. Of the different concentrations of priming agents tested, the effect of a moderate concentration of kinetin （150 mg/L） was very pronounced, particularly in improving growth and grain yield, in both cultivars. In addition, priming with kinetin alleviated the adverse effect of salt stress on gaseous exchange characteristics （net CO2 assimilation rate and water use efficiency） in both cultivars. Seed priming with a moderate concentration of kinetin also altered the pattern of accumulation of Na^＋ and Clˉ in the shoots, irrespective of the wheat cultivar, under conditions of salt stress. However, all other priming agents at the different concentrations tested did not show any consistent effect on ion levels, except hydropriming, which increased K^＋ levels in the shoots of both cultivars under salt stress. In conclusion, a moderate concentration of kinetin showed a consistent effect in altering the growth and grain yield of both wheat cultivars, which was related to the beneficial effects of kinetin priming on water use efficiency and photosynthetic rate under conditions of salt stress.     

Effects of Phosphorus Nutrient on the Hydraulic Conductivity of Sorghum （Sorghum vulgare Pers.） Seedling Roots Under Water Deficiency

Hydroponic experiments were conducted in a growth chamber and changes in the hydraulic conductivity of sorghum (Sorghum vulgare Pers.) roots (Lpr) at the three-leaf stage were measured using the pressure chamber method. Water deficiency was imposed with polyethylene glycol (PEG) 6000 and the phosphorus (P) levels were controlled by complete Hoagland solution with and without P nutrient. The objective of this study was to investigate the effect of P nutrition on root Lpr under water deficiency. The results showed that the Lpr in P deficiency treatments decreased markedly, but the Lpr recovered to the same value as that of control when sufficient P was supplied for 4-24 h. Water deficiency decreased Lpr, but the hydraulic conductivity of the roots with sufficient P supply was still higher than that of plants without P supply. When resuming water supply, the Lpr of the water-deficient plants under P supply recovered faster than that of plants without P supply, which indicates that plants with sufficient P nutrient are more drought tolerant and have a greater ability to recover after drought. The treatment of HgCl2 indicated that P nutrient could regulate the Lpr by affecting the activity and the expression levels of aquaporins.     

Antiporter Gene from Hordum brevisubulatum （Trin.） Link and Its Overexpression in Transgenic Tobaccos

A vacuolar Na^ /H^ antiporter cDNA gene was successfully isolated fromHordeum brevisubulatum (Trin.) Link using the rapid amplification ofcDNA ends (RACE) method. The gene was named HbNHXI and was found to consist of 1 916 bp encoding a predicted polypeptide of 540 amino acids with a conserved amiloride-binding domain. Phylogenetic tree analysis of the Na^ /H^ antiporters showed that the HbNHXI gene shares 55.3%-74.8% similarity with the vacuolar-type Na^ /H^ antiporters. Transgenic tobaccos that contain the HbNHXI gene, integrated by forward insertion into the tobacco genome, were obtained via Agrobacterium tumerfaciens and characterized for the determination of the concentration of Na^ and K^ ions, as well as proline, in the presence of 300 mmol/L NaCl. The T1 transgenic plants showed more tolerance to salt and drought than did wild-type plants. Our data suggest that overexpression of the HbNHXI gene could improve the tolerance of transgenic tobaccos to salt and drought through the function of the vacuolar Na^ /H^ antiporter.     

拟南芥MT-II过量表达提高抗旱性(英文)

富含巯基的植物II型金属硫蛋白(MT)对植物抵抗重金属胁迫具有重要作用,其中一个可能机制是金属硫蛋白可能猝灭重金属引起的氧化胁迫。利用转MT-II基因和野生型拟南芥(Arabidopsis thaliana)植株来对比研究MT在胁迫过程中通过清除氧自由基,特别是H2O2而对植物抗旱性的影响。研究表明,转基因型拟南芥能有效维持体内氧化—还原势,减少MDA的产生,从而缓解干旱胁迫引起的伤害,提高抗旱性。     

扣囊复膜孢酵母β-葡萄糖苷酶基因在毕赤酵母中的克隆与表达

利用PCR技术,从扣囊复膜孢酵母的总DNA中扩增得到β-葡萄糖苷酶（β-Glucosidase）基因 (BGL1),长度为2596 bp,连接到pGEMT载体上,用限制性内切酶切下目的基因,插入到巴斯德毕赤酵母表达载体pPIC9K中,使之位于α-因子信号肽下游,且与之同框, 构建成重组质粒pSHL9K。 通过电转化将重组质粒pSHL9K插入到Pichia pastoris GS115菌株染色体中,获得高效表达BGL1基因的毕赤酵母重组工程菌株。重组酶的最适温度为50℃,最适pH为5.4。培养基中β-葡萄糖苷酶活性最高可达47U/mL。     

外源β-胡萝卜素、光照对青霉PT95菌株菌核分化和类胡萝卜素产率的影响

初步研究了外源β-胡萝卜素和光照对青霉PT95菌株菌核分化和类胡萝卜素产率的影响。结果表明,在培养基中加入外源β-胡萝卜素后,PT95菌株渗出液出现的时间、菌核出现的时间延迟了,但菌核成熟的时间没变。培养基中的外源β胡萝卜素浓度越大,其渗出液、菌核出现的时间越迟。外源β胡萝卜素亦能降低PT95菌株的脂质过氧化水平和菌核中的类胡萝卜素含量。高氧胁迫的光照培养条件有利于PT95菌株的菌核分化和色素在菌核中的积累;与低氧胁迫的黑暗培养条件相比,其菌核生物量和类胡萝卜素产率分别增加了18.7%和101%。以上实验结果表明,若想获得高的菌核生物量和类胡萝卜素产率,应该尽可能在高氧胁迫、无抗氧化剂存在的条件下培养PT95菌株。     

鼠伤寒沙门氏菌SL7207 SifA-突变株的构建和鉴定

鼠伤寒沙门氏菌SifA^-基因突变株的特点是能进入真核细胞的胞液。利用P22噬菌体转导技术构建了鼠伤寒沙门氏菌疫苗株SL7207的SifA^-突变株SL7207,该突变株与SL7207有着相似的体外生长曲线和细胞侵袭力,SL7207。在MDCK上皮细胞中的增殖能力增强,但在RAW264．7巨噬细胞中的生存能力减弱。小鼠毒力试验显示SL7207。在BALB／c小鼠体内毒力下降。仅SL7207在体外可向RAW264．7巨噬细胞递送真核表达质粒。SL7207的构建为重组沙门氏菌疫苗载体的研制提供了一个新的选择。