中国野生葡萄抗霜霉病相关基因VpPR4b及其启动子的克隆和功能分析

以中国野生葡萄(Vitis piasezkii)‘留坝-8’(抗霜霉病)和欧亚种葡萄(V.vinifera)‘黑比诺’(易感霜霉病)为材料,克隆得到抗霜霉病相关基因VpPR4b和VvPR4b。序列分析发现VpPR4b和VvPR4b核苷酸序列相似性为98.61%。VpPR4b编码区为432 bp,编码143个氨基酸,含有BARWIN保守结构域,属于典型的Ⅱ型PR4蛋白。VpPR4b和VvPR4b均可在霜霉菌侵染后诱导表达。亚细胞定位表明VpPR4b定位于细胞膜上。分别从‘留坝-8’和‘黑比诺’中克隆得到了VpPR4b和VvPR4b的启动子,其序列相似性高达95.65%,并且含有相似的顺式作用元件。通过酵母单杂交验证了转录因子WRKY40和WRKY75可以结合VpPR4b启动子。本研究的结果表明PR4b可能在葡萄抵御霜霉病侵染过程中发挥一定作用。     

Defense response of tomato fruit at different maturity stages to salicylic acid and ethephon

In order to elucidate whether fruit maturity stage influence the induced resistance of exogenous elicitors in tomato and the involved mechanisms, we investigated the defense responses of tomato fruits against Botrytis cinerea, ethylene production and internal quality following treatments of fruit with salicylic acid (SA) or ethephon (ET) at mature green (MG) and breaker (BR). SA significantly suppressed decay and disease incidence in tomato fruits at both MG and BR stages, along with higher expression level of PR1 gene after 2 days of treatment. All fruits treated by SA had lower contents of ethylene and lycopene. The ET-treated fruit at both maturity stages showed lower disease incidence and higher level of PR2 and PR3 expression compared with the control fruit. ET treatment significantly enhanced ethylene and lycopene contents, and accelerated fruit ripening. Our results suggest that SA and ET induced disease resistance in fruits by mediating the expression of different pathogenesis-related genes and have different effects on fruit ripening, which in turn influences the disease resistance of tomato fruits.     

Hydro-physical responses of gypseous and non-gypseous soils to livestock grazing in a semi-arid region of NE Spain

Pasture productivity depends on soil hydro-physical properties, which in turn are deeply affected by livestock grazing. However, the comparative response of different soil types, and particularly gypseous soil types, to grazing has hardly been studied before. This paper compares the effect of grazing on the soil hydro-physical properties of silty gypseous (Gy) and non-gypseous (NGy) soils located in a semi-arid region (Middle Ebro Valley, NE, Spain). Two different soil managements were selected: ungrazed natural shrubland (N) and grazed shrubland (GR) soils. The gypsum, CaCO₃ and organic matter content (OM), soil texture, soil bulk density (ρ_b), penetration resistance (PR), saturated sorptivity (S), hydraulic conductivity (K), and the water retention curve (WRC) for undisturbed soil samples from 1 to 10 cm depth soil layer were measured. The ρ_b and PR in NGy soils were significantly higher than those observed in the Gy ones. Soil compaction due to grazing treatment tended to increase ρ_b and decrease the K and S values. While no differences in PR were observed in the Gy soils between grazing treatments, the PR measured in the NGy soils under GR was significantly higher than the corresponding values observed under N. Differences in K and S between GR and N treatments were only significant (p < 0.05) in NGy soils, where K and S values under the N treatment were almost four times greater than the corresponding values measured under GR. Overall, no differences in the WRCs were observed between soil types and grazing treatments. While the WRCs of NGy soils were not significantly affected by the grazing treatment, Gy soils under N treatment present a significantly higher level of soil macropores than under GR treatment. The hydro-physical features of Gy soils tended to be less affected by grazing than those of the NGy soils. These results suggest that livestock grazing, in both Gy and NGy soils, has a negative effect on the physical soil properties, which should be taken into account by land managers of these semi-arid regions where silty gypseous and non-gypseous areas coexist.     

2010年上半年11省市规模猪场疫病监测与剖析

为了解近期发病猪场几种病毒性疫病的流行状况,采用RT-PCR和PCR方法,对2010年1月至6月期间,来自11省(市) 50个猪场共147份病料进行猪瘟病毒（CSFV）、猪繁殖与呼吸综合征病毒（PRRSV）、高致病性猪繁殖与呼吸综合征病毒（HP-PRRSV）、伪狂犬病毒（PRV）和猪圆环病毒2型（PCV2）检测,并采用ELISA方法对其中26个猪场458份血清样品进行伪狂犬gE野毒抗体检测。结果表明,PCV2阳性率最高,为75.3%（70/93）;PRRSV阳性率次之,为57.4%（78/136）,HP-PRRSV阳性率为43.5%（54/124）;CSFV和PRV的阳性率分别为37.7%（52/138）和21.0%（21/100）;伪狂犬gE野毒抗体阳性率为16.4%（75/458）。同时检出2种及2种以上病毒的样品占46.9%（69/147）,其中以HP-PRRSV+PCV2、CSFV+PCV2及CSFV+HP-PRRSV+PCV2三种形式最为常见。PRRSV（50.0%）是猪场繁殖障碍性疾病中的最常见病原;CSFV（56.0%）和PCV2（48.0%）是断奶前后仔猪腹泻的常见病原;HP-PRRSV（53.5%）、CSFV（50.7%）和PCV2（48.0%）是保育猪发生高热症的常见病原。该检测结果为猪场疫病的诊断与防控提供了参考。     

烟草PR10蛋白生物活性及赤星病菌Alternaria alternata诱导下的表达分析

为深入研究植物PR10蛋白的生物学功能,以烟草PR10蛋白(NtPR10)为研究对象,采用冷休克蛋白表达载体p Cold II,构建烟草PR10融合蛋白原核表达系统,优化表达及纯化条件,获得高纯度NtPR10融合蛋白;分别采用底物法和滤纸片法体外分析其核酸酶活性及抑菌活性;并利用荧光定量PCR方法分析了烟草赤星病菌Alternaria alternata侵染下,NtPR10基因在抗、感烟草品种内不同时间点的表达差异。结果表明,15℃、0.1 mmol/L IPTG过夜条件下可诱导获得大量可溶性目的蛋白,50、100 mmol/L咪唑缓冲液洗脱能够获得较高纯度的目的蛋白;纯化后的NtPR10蛋白能够降解烟草总RNA,具有核酸酶活性,且1.0、0.5、0.25μg/μL的NtPR10蛋白溶液均对烟草赤星病菌的菌丝生长具有显著的抑制作用,但随着蛋白浓度的降低,抑菌作用减弱;荧光定量PCR结果显示,接种烟草赤星病菌后,NtPR10基因在感病品种和抗病品种中均显著上调表达,但其在抗病品种的响应速度和表达量显著高于感病品种,表明NtPR10应答了烟草赤星病菌的侵染过程。     

雌激素和孕酮对犬子宫内膜基质细胞增殖和孕酮受体表达的影响

本试验应用不同消化分离途径获取犬子宫内膜基质细胞,调节培养液中雌激素(E₂)和孕酮(P₄)的浓度,采用MTT法测定E₂和P₄浓度水平对犬子宫内膜基质细胞体外增殖的影响,利用细胞免疫组织化学法鉴定细胞并测定细胞孕酮受体(PR)表达与激素浓度水平的相关性。结果表明,E₂浓度变化(15、30、100 pg/mL)对犬子宫内膜基质细胞的增殖和PR的表达均没有显著的调节作用(P>0.05);P₄(15、30 ng/mL)对犬子宫内膜基质细胞的增殖有显著促进作用(P<0.05),P₄(3、15、30 ng/mL)对犬子宫内膜基质细胞PR的表达具有显著的抑制作用(P<0.05),其影响程度与浓度和作用时间关系密切。     

巴西橡胶树PR107生物量研究初探

利用橡胶树生物量估测数学模型,测算了海南儋州地区2～28龄巴西橡胶无性系PR107生物量的年度变化与累积量,结果表明：（1）住橡胶树生命周期内,茎围与生物量的增长量呈线性增加,并在开割前期出现高峰期,在开割后逐渐变缓慢,进入老龄时,生长速度更慢;橡胶树茎围生长受环境的影响较明显,与树龄、季节、月平均气温、月降雨量和月日照时数直线回归达极显著水平,其回归方程为：y=0．062380—0．013408x,＋0．008805x2＋0．015716x3＋0．000231x4＋0．000002379x5;（2）橡胶树各器官（叶、枝、干、皮、根）干物质积累量不仅与季节有关,还与树龄有关。随着一年中季节的变化,干物质积累量呈增长趋势;而随着树龄的增长,枝、干、皮干物质积累量呈增长趋势,而根和叶则呈下降趋势;（3）橡胶树生物量受环境因素和人为因素的影响。橡胶树生物量增量与树龄、季节、月平均气温、月降雨量和月日照时数直线回归达极显著水平,其回归方程为：Y=-1661．788621—307．824821x1＋209．482331x2＋327．130313x3＋7．737860x4—9．343308x5;（4）橡胶树生物量的分配格局因不同年龄和器官而异。树干生物量最大,其次是小枝,而其它器官生物量则因年龄大小而有所变化。但在橡胶树整个生命周期内,各器官生物量大小顺序为：树叶〉树干〉干胶〉树根〉树皮。     

橡胶人工林生态系统养分循环研究I．无性系PR107氮素体循环

通过对儋州地区,林分结构相近、立地条件相似、抚管措施相同、林龄不同（2a～28a）的橡胶无性系PR107的生物量和分配格局、生产力、氮素积累与固定、氮素归还、凋落物及氮素利用效率进行比较研究,发现它们之间存在较明显的差别,并表现出不同的养分生物学特性。试验结果表明：（1）橡胶树不同器官氮素含量高低顺序为：叶>树枝>树根>树皮>树干>胶乳,而积累量和分配格局则为：树干>树枝>树叶>树皮>树根>胶乳。在橡胶树28a生长周期中,养分积累总量可达5639.12kg/hm2,其中氮素约为1606.13kg/hm2,占28.48%。（2）随着树龄的增长,橡胶树枯落物量和由此向土壤归还的氮素总量也逐年增加,氮素归还率也逐年增加。据测定,不同树龄橡胶树每年枯落物总量约为0.295～11.08t/hm2,由枯落物向土壤归还的养分总量达13.44～431.4kg/hm2.a,其中氮约为5.28～162.62kg/hm2。（3）橡胶树体对氮素的利用效率基本上随着树龄的增加逐渐降低,各器官对氮素的利用效率大小顺序为：树干>树皮>胶乳>树枝>树根>树叶。     

Agrobiodiversity loss and livelihood vulnerability as a consequence of converting from subsistence farming systems to commercial plantation‐dominated systems in Xishuangbanna,Yunnan, China: A household level analysis

Production of rice (Oryza sativa) under a shifting cultivation system (swidden rice) and paddy fields (wet rice) is an important livelihood strategy in Xishuangbanna, Yunnan, China where households are facing conversion from subsistence systems to commercial plantation‐dominated systems represented by smallholder rubber plantations. However, there are few studies of the effects on rice production and varieties diversity, as well as livelihood effects at a household level during this conversion. This paper presents a household level analysis of agrobiodiversity loss and food security, as well as livelihood vulnerability during agroecosystems' conversion in Xishuangbanna. First, a quantitative assessment was made of the statistical data for rice and rubber production during the past 50 years in Xishuangbanna and at a village level from 1985 to 2005 for three villages from the valley floor and upland hills, respectively. Second, at a household level, 60 per cent of households from the three villages were investigated with a questionnaire from 1998 to 2005. The process of transformation has led to significant changes in the household's income and its proportion. Meanwhile, households have suffered livelihood vulnerability from excessive rubber plantations. The centrality of rice in Southeast Asian agricultural and social systems, contrasted with the significant erosion of rice‐based agroecosystem of local households, calls for increased attention to the livelihood vulnerability during this conversion. Correspondingly, practical strategies to resolve livelihood vulnerability are proposed, such as construction of paddy and effective irrigation systems for food security, agroforestry systems in rubber plantation, instead of monoculture, and diversification of sources of household income. Copyright © 2010 John Wiley & Sons, Ltd.