Agrobacterium‐produced and exogenous cytokinin‐modulated Agrobacterium‐mediated plant transformation

Agrobacterium tumefaciens is a plant pathogenic bacterium that causes neoplastic growths, called ‘crown gall’, via the transfer and integration of transferred DNA (T‐DNA) from the bacterium into the plant genome. We characterized an acetosyringone (AS)‐induced tumour‐inducing (Ti) plasmid gene, tzs (trans‐zeatin synthesizing), that is responsible for the synthesis of the plant hormone cytokinin in nopaline‐type A. tumefaciens strains. The loss of Tzs protein expression and trans‐zeatin secretions by the tzs frameshift (tzs‐fs) mutant is associated with reduced tumorigenesis efficiency on white radish stems and reduced transformation efficiencies on Arabidopsis roots. Complementation of the tzs‐fs mutant with a wild‐type tzs gene restored wild‐type levels of trans‐zeatin secretions and transformation efficiencies. Exogenous application of cytokinin during infection increased the transient transformation efficiency of Arabidopsis roots infected by strains lacking Tzs, which suggests that the lower transformation efficiency resulted from the lack of Agrobacterium‐produced cytokinin. Interestingly, although the tzs‐fs mutant displayed reduced tumorigenesis efficiency on several tested plants, the loss of Tzs enhanced tumorigenesis efficiencies on green pepper and cowpea. These data strongly suggest that Tzs, by synthesizing trans‐zeatin at early stage(s) of the infection process, modulates plant transformation efficiency by A. tumefaciens.     

丁(鱼岁)不同群体间形态学差异与随机扩增多态DNA(RAPD)分析

采用聚类分析、主成分分析和判别分析3种数理统计方法,对我国新疆朱家湖丁(鱼岁)群体、73水库丁(鱼岁)群体和从捷克引进我国的丁(鱼岁)群体的可量性状和框架参数进行分析.聚类分析和主成分分析显示,朱家湖群体与73水库群体较为接近,而捷克群体与前两群体相距较远.判别分析亦可将捷克群体与前两群体分开,准确率达100%.从60个随机引物中筛选出的20个引物对丁(鱼岁)朱家湖群体、73水库群体和捷克群体进行了随机扩增多态DNA(RAPD)分析.引物S440在捷克群体扩增出的450bp片段为该群体特有的标记片段.朱家湖群体、73水库群体、捷克群体多态位点比例分别为24%、22.67%和18.42%;群体内遗传相似度分别为0.8967、0.9035和0.9309;遗传多态度(π)分别为0.1539、0.1489和0.1142.表明朱家湖群体保持着较高的遗传变异.三群体间的遗传相似度为0.6868-0.9496,群体遗传分化系数(Fst)为0.048-0.238,分子方差分析发现群体内方差占总方差的83.96%,群体间的方差只占16.04%,由此推断三群体间遗传分化并不大.     

植物源调节剂对水稻的影响

植物源调节剂是利用艾蒿为主要原料,首次用提取的混合物研制成的一种植物生长调节剂,使用80mg/L的植物源调节剂混合物水溶液,分别在水稻苗期、拔节期、孕穗期叶面喷洒,清水对照。结果表明:植物源调节剂使水稻增产11.38%,千粒重增加11.45%。株干重增加了17.1%。在灌浆期连续测定表明:相对于CK,穗粒重增加10.9%,叶绿素含量提高14.41%。品质分析结果为:可溶性糖含量提高15.57%、游离氨基酸含量提高18.52%、淀粉含量提高1.59%,可溶性蛋白质降低22.17%、粗纤维降低1.95%、粗脂肪降低1.22%。该植物源调节剂对水稻的生长和产量都有显著的促进效应。     

螺旋藻转化纳米元素硒的制备及其体外清除自由基活性的初步研究

研究利用高密度富硒螺旋藻(Se-SP)细胞通过生物转化制备纳米元素硒(Nano-Se)的可行性,观察Nano-Se在体外对氧自由基的清除作用。用梯度离心分选Nano-Se,原子力显微镜(AFM)、透射电镜(TEM)及X-射线能谱(EDX)联用表征纳米粒中的元素硒形态,电感耦合等离子质谱仪(ICP-MS)测定Nano-Se中的硒含量,化学发光方法检测Nano-Se在体外对超氧自由基和羟自由基的清除作用。结果发现,Nano-Se主要由元素硒构成,形态呈球形,73%的纳米粒子直径大小分布在(61±17)nm范围。Nano-Se在体外对两种氧自由基的最大清除率分别为:30.1%和27.6%,相应的EC50分别为:0.8 μg/ml和2.2 μg/ml。相同剂量时,Nano-Se对氧自由基的清除作用比硒代蛋氨酸(Se-Met)及Se-SP中其它含硒活性成分更强。结果提示,利用高密度Se-SP可诱导Nano-Se的大量生成,Se-SP转化的Nano-Se可能是一种新的抗氧化硒形态,其作用机制和体内生物活性有待深入研究。     

Phospholipase Dδ is Involved in Wounding Induced Phosphatidic Acid Formation in Arabidopsis

Phosphalipase D (PLD) hydrolyzes phospholipids into phosphatidic acid (PA). PLDα1 and δ are the two most abundant members of the 12 member PLD family in Arabidopsis. PLDα1 has been demonstrated having role in the wounding induced PA signalling. However, whether and how PLDδ is involved in wounding induced PA formation remained unclear. In the present study, the membrane lipids response to wounding was profiled in Wassilewskija (WS) and PLDδ knockout mutant (PLDδ KO) of Arabidopsis. The levels of most lipids, including monogalactosyldiacylglycerol, digalactosyldiacylglycerol, phosphatidylcholine and phosphatidylglycerol had decreased rapidly within 30min after wounding in the two Arabidopsis genotypes. In contrast, the level of PA increased sharply and significantly 30min after wounding. It continued to increase until peaking at 1h post wounding in WS and 3h post wounding in PLDδ KO, and then decreased. The PA levels were similar in the two genotypes in untreated leaves and in leaves 6h after wounding. However, these levels were lower in PLDδ KO than in WS from 30min to 3h post wounding. The significant difference of PA level between the two genotypes occurred 30min after wounding, when it was about 20% lower in PLDδ KO than in WS. These results show that PLDδ is involved in wounding induced PA formation in Arabidopsis, but its absence induces PA response later and with less intensity than PLDα1.     

陕西凤翔孙家南头秦墓出土人骨中C和N同位素分析

本文对陕西凤翔孙家南头秦墓出土的春秋战国时期人骨中C和N稳定同位素组成进行了测试分析。结果表明: 人骨样品均可用于食谱分析; 由δ13C和δ15N值综合分析可知, 春秋战国时期陕西凤翔地区秦先民是以C4类植物为主食、辅以少量肉食的杂食性食谱; 经济模式可能为农业与畜牧兼营, 以种植黍、粟类等旱地作物的农业形式为主, 并辅以驯养牲畜; 根据墓主与殉人的δ13C和δ15N值对比分析, 推测殉人可能是与墓主饮食方式较为相近的姬妾或仆从之类的人;春 秋中期至战国晚期秦先民的食物结构较为稳定, C4植物比例与δ15N发生变化的原因可能与当时历史的变迁与社会的发展有一定的关系。     

HCV丝氨酸蛋白酶小分子底物构建及GST融合表达

丝氨酸蛋白酶是丙型肝炎病毒重要的功能蛋白和药物作用靶点,其通过分子内（cis）和分子间（trans）方式催化水解前体蛋白,释放病毒功能蛋白。目的：为深入研究病毒蛋白酶活性和抑制剂鉴定需要,实验研究参照丙型肝炎病毒1a亚型菌株蛋白酶天然底物的氨基酸序列特点,设计了一段包含两个天然底物酶切位点的小分子多肽2S,并进行了原核表达。方法：利用PCR方法,合成2S小分子多肽基因,目的基因两端引入BamH I和EcoR I两个限制性酶切位点,双酶切后将基因与表达载体pGEX-4T-2重组,转化大肠杆菌DH5α,经化学诱导进行GST融合蛋白表达,通过亲和层析柱纯化目的蛋白。纯化的GST 2S融合蛋白在体外反应系统进行酶切鉴定,SDS-PAGE和ELISA鉴定酶切结果。结果：PCR合成的小分子底物多肽2S基因,经与表达载体重组后测序,证实基因序列正确。采用0.5mmol/L浓度的IPTG诱导工程菌过夜,获得表达的目的蛋白,经分离纯化得到融合蛋白GST-2S。GST-2S在体外磷酸盐缓冲系统中与丝氨酸蛋白酶反应,15%SDS-PAGE鉴定酶切产物,证实融合蛋白底物条带明显消失,ELISA结果同样说明融合蛋白的底物活性。结论：含有两个天然底物酶切位点的小分子多肽可以替代病毒天然底物,实验结果为丙型肝炎病毒丝氨酸蛋白酶活性研究和酶抑制剂研究奠定了方法学基础。     

四川老君山自然保护区红翅噪鹛冬季栖息地特征

摘要：2010年1月和2011年1月,在四川老君山国家级自然保护区对红翅噪鹛（Garrulax formosus）的冬季栖息地特征进行了初步研究。在研究区内,红翅噪鹛冬季多集小群活动,主要选择次生林,而回避原生林和人工林。2个冬季在野外共遇见红翅噪鹛21群78只。通过比较分析,发现红翅噪鹛喜欢在海拔较低、坡位较高、坡向...     

中国和越南褐飞虱抗药性研究

2010年用稻茎浸渍法监测了我国和越南共14个褐飞虱Nilaparvata lugens(St(a)l)田间种群对5种杀虫剂的抗性,结果表明:14个褐飞虱田间种群对吡虫啉、噻嗪酮、氟虫腈、吡蚜酮和叶蝉散的LC50值分别在9.5287～46 6716、1.6621～17.8785、0.9818～ 8.4084、0.331...     

Lacking chloroplasts in guard cells of crumpled leaf attenuates stomatal opening: both guard cell chloroplasts and mesophyll contribute to guard cell ATP levels

Controversies regarding the function of guard cell chloroplasts and the contribution of mesophyll in stomatal movements have persisted for several decades. Here, by comparing the stomatal opening of guard cells with (crl‐ch) or without chloroplasts (crl‐no ch) in one epidermis of crl (crumpled leaf) mutant in Arabidopsis, we showed that stomatal apertures of crl‐no ch were approximately 65–70% those of crl‐ch and approximately 50–60% those of wild type. The weakened stomatal opening in crl‐no ch could be partially restored by imposing lower extracellular pH. Correspondingly, the external pH changes and K⁺ accumulations following fusicoccin (FC) treatment were greatly reduced in the guard cells of crl‐no ch compared with crl‐ch and wild type. Determination of the relative ATP levels in individual cells showed that crl‐no ch guard cells contained considerably lower levels of ATP than did crl‐ch and wild type after 2 h of white light illumination. In addition, guard cell ATP levels were lower in the epidermis than in leaves, which is consistent with the observed weaker stomatal opening response to white light in the epidermis than in leaves. These results provide evidence that both guard cell chloroplasts and mesophyll contribute to the ATP source for H⁺ extrusion by guard cells.