大麦与油菜种皮特异启动子表达模式的比较

启动子位于转录起始位点上游并能特异性地结合RNA聚合酶,其作为调控序列驱动外源基因在异源植物中表达,从而实现转基因的高效性,具有时空表达特异性的启动子对获得有效转基因植物及产物具有重要意义。为了解种皮特异启动子的表达模式,该研究基于前期报道的序列,通过同源克隆的方法分别从大麦和油菜中克隆获得Gerb和Bntt两个种皮特异性启动子,并对其进行生物信息学分析,构建了Gerb::GUS和Bntt::GUS植物表达载体并转化拟南芥,通过组织化学染色观察了GUS的表达情况。结果表明:两种启动子序列中都含有多拷贝种皮特异表达启动子元件以及多种胁迫诱导响应元件;转基因拟南芥幼苗期,大麦Gerb种皮特异启动子驱动GUS全株表达且子叶和下胚轴较真叶和根中表达量高;油菜Bntt种皮特异启动子表达较弱;成株期,Gerb在不同组织(叶片、茎、花序和角果)中均有表达,未显示组织特异性;Bntt仅在叶片及角果维管束中有微弱表达。在各种非生物胁迫下,Gerb表达模式未发生显著变化,而Bntt仅在盐胁迫下显示很强的角果和种子特异性表达,其他胁迫未见明显表达。以上结果显示,大麦种皮特异性启动子Gerb和油菜种皮特异性启动子Bntt在时间和空间表达模式上存在差异,这对今后选择种皮特异启动子具有参考作用,但其具体机制仍需进一步研究验证。     

低温胁迫下枇杷幼果种子膜脂过氧化、保护酶活性及显微结构的变化

枇杷(Eriobotrya japonica)开花结果正值冬季低温期,易受低温影响,因此寒害成为抑制枇杷健康生长、发育的重要因素之一。种子作为产生多种内源激素的中心,其健康程度与枇杷果实正常发育息息相关。该研究以四川省成都市龙泉驿区柏合镇的‘早钟六号’枇杷幼果种子为材料,经不同低温(6、3、0、-3℃)胁迫不同时间(12、24、36、48 h)后,对其相对电导率、丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和过氧化物酶(POD)的变化进行测定以及细胞解剖结构分析,旨在探究枇杷幼果种子抗寒机制,为提高四川地区枇杷抗寒栽培新技术提供理论和实践依据。结果表明:低温胁迫下,枇杷幼果种子相对电导率及MDA含量随着处理温度的降低整体呈上升趋势;保护酶SOD、CAT活性在低温胁迫前期不同程度升高,至某个低温后呈下降趋势;而POD活性总体呈升-降-升趋势;相对电导率、MDA含量、SOD活性、CAT活性、POD活性的变化临界温度均为6℃,胁迫临界时间分别为12、24、48、36 h,而MDA含量变化临界温度为3℃,胁迫临界时间为36 h;显微结构表明枇杷幼果种子6℃低温开始受冻,最先受冻部位为种皮,其次为种胚真叶原始体或真叶,最后到细胞破裂,说明枇杷幼果种子随着处理温度降低,细胞结构受损越严重。综上,枇杷幼果种子受冻临界温度为6℃,受冻临界时间为12 h。     

盐胁迫对低温预处理石榴种子萌发及幼苗生理生化的影响

以催芽‘峄城大青皮甜’石榴种子为材料,经2~4℃低温预处理5、10和15d,置于不同盐分梯度下进行种子萌发试验,并测定分析幼苗生长、细胞保护酶活性、可溶性糖和脯氨酸含量及电导率变化,以阐明盐胁迫下低温预处理对石榴种子萌发和幼苗生长及生理生化的影响。结果显示:(1)适当的低温预处理和低盐胁迫能提高石榴种子的发芽率和发芽势,低温处理5d和0.1%低盐环境下有利于壮苗生长。(2)适当时间的低温预处理(5d)可显著提高石榴幼苗的保护酶(SOD、POD、CAT)活性,长时间的低温预处理(15d)却降低了幼苗的保护酶活性;随着盐浓度的升高石榴幼苗的保护酶活性也逐渐升高,并在盐浓度为0.3%时均达到最大值,说明当盐浓度高于0.3%时超出了石榴幼苗保护酶系统的调节范围,从而发生盐害。(3)低温预处理5d和10d降低了幼苗可溶性蛋白和脯氨酸的含量;随着盐浓度的升高幼苗可溶性蛋白和脯氨酸的含量也逐渐升高。(4)5d和10d的低温预处理可以提高石榴幼苗细胞膜透性,随着盐浓度的升高相对电导率也显著升高。研究表明,在低盐环境下,适当时间的低温预处理能显著促进石榴种子萌发生长,其幼苗能通过调整自身渗透调节物质可溶性蛋白和脯氨酸含量以及体内保护酶活性,降低细胞膜脂过氧化程度,保证细胞膜的完整性,维持正常的细胞代谢活性,从而有效减轻盐胁迫对植株的伤害,表现出较强的抗盐性。     

棉花纤维发育相关基因GhPRP10的克隆及其功能分析

利用电子序列拼接结合RT-PCR技术,从12DPA(开花后天数)棉纤维中克隆到1个编码富含脯氨酸蛋白(PRPs)基因,命名为GhPRP10(登录号KP036633)。GhPRP10基因开放阅读框为684bp,编码228个氨基酸,其中脯氨酸(Pro)含量为34.6%。序列分析发现GhPRP10蛋白具有N端信号肽和富含脯氨酸区域,属于第一类PRPs。实时荧光定量PCR(RT-PCR)结果显示,GhPRP10在棉纤维组织中优势表达,在纤维发育过程中的表达量呈现先升高后降低的趋势,在18DPA纤维中表达量最高。利用Gateway技术构建植物过量表达载体,转入烟草BY-2悬浮细胞,表型观察和细胞长度测量结果显示,转GhPRP10基因细胞比野生型细胞显著增长。根据该基因的组织表达特征和转基因细胞表型分析,推测GhPRP10基因在纤维伸长和次生壁合成过程中发挥作用。     

昼夜温差对番茄生长发育、产量及果实品质的影响

在3间人工气候室精密受控环境中,保持日平均温度为22 ℃,设置昼夜温差分别为6 ℃(25 ℃/19 ℃)、8 ℃(26 ℃/18 ℃)、10 ℃(27 ℃/17 ℃),研究昼夜温差对番茄生长的影响.结果表明: 番茄不同品种、不同生长时期适宜的昼夜温差条件不同.番茄开花前,与6 ℃温差相比,8 ℃温差可显著加快野生种醋栗番茄LA1781生长发育,使幼苗株高增加23.1%,出叶加快1～2片,开花提前7 d;10 ℃温差对LA1781苗期的促进作用与8 ℃温差相似.对栽培种普通番茄LA2397和LA0490来说,6 ℃温差使幼苗生长良好,8 ℃温差对幼苗无显著促进作用;与6 ℃温差相比,10 ℃温差对苗期生长及开花有抑制作用,使株高降低12.0%～18.3%,出叶慢2～3片,开花推迟2～4 d.10 ℃温差使3个品种番茄地上部分干质量增加25.2%～44.2%.番茄开花后,与6 ℃温差相比,10 ℃温差可显著提高LA1781的产量和果实品质,使果实数增加34.7%,单株产量增加92.1%,平均单果质量增加40.0%,果实可溶性糖含量增加16.3%,番茄红素含量增加95.6%.与6 ℃温差相比,LA2397和LA0490在8 ℃温差下产量和果实品质提高,番茄红素含量增加超过2倍;在10 ℃温差下产量略有降低(5.0%),果实含糖量降低,但果实尺寸和番茄红素含量增加.表明番茄苗期生长温差不宜过大,花果期适当增大昼夜温差可提高产量和果实品质,但温差过大易造成生长不良和减产.     

高温胁迫对Bt棉铃壳中Bt蛋白含量及氮代谢的影响

以Bt基因来源于中国的棉花品种泗抗1 号(常规种)、泗抗3 号(杂交种)和来源于美国的棉花品种99B(常规棉)、岱杂1 号(杂交棉)为材料,研究了不同高温水平下Bt 棉盛铃期铃壳中Bt 蛋白含量变化及氮代谢生理特征.结果表明: 铃壳中Bt 蛋白含量随温度升高而降低,与对照相比(32 ℃),常规棉品种在38 ℃、杂交棉品种在40 ℃以上时,铃壳中Bt 蛋白含量大幅度下降.其中,常规种泗抗1号和99B在38 ℃时分别下降53.0%和69.5%;杂交种泗抗3号和岱杂1号在40 ℃时下降64.8%和54.1%.铃壳Bt 杀虫蛋白含量下降显著时,其可溶性蛋白含量明显下降,游离氨基酸含量明显提高,GPT活性显著下降,蛋白酶活性显著增加.高温影响铃壳的氮代谢引起Bt蛋白的分解加剧,合成减弱,从而造成Bt蛋白含量减少,抗虫性下降.     

Tracing dietary origins of aphids and the predatory beetle Propylea japonica in agricultural systems using stable isotope analyses

Tracing dietary origins of the predatory beetle Propylea japonica (Thunberg) (Coleoptera: Coccinellidae) aids understanding their roles in the food web and provides information to develop strategies for effective conservation in agroecosystems comprised of wheat [Triticum aestivum L. (Poaceae)], cotton [Hirsutum spp. (Malvaceae)], and maize [Zea mays L. (Poaceae)]. Intrinsic markers of carbon and nitrogen stable isotope ratios (δ¹³C and δ¹⁵N) in P. japonica need to be developed to ascertain the source(s) of diet. Experiments were carried out to examine the changes of δ¹³C and δ¹⁵N among the three crops, pests (wheat, cotton, and maize aphids; all Hemiptera: Aphididae), and P. japonica fed on aphids of each of the three crops. Results indicated that δ¹³C values in P. japonica fed on wheat, cotton, and maize aphids were ?27.2 to ?26.5‰, ?24.2 to ?23.9‰, and ?11.0 to ?10.7‰, respectively, whereas their δ¹⁵N values were 1.1 to 2.9‰, 6.0 to 7.4‰, and ?0.6 to 0.1‰, respectively. δ¹³C and δ¹⁵N plots clearly identify the three crops, the dietary origins of the aphids, and the host origins of the aphid prey consumed by the ladybird beetles, as each pathway displays a non‐overlapping pattern. Based on the values of δ¹³C and δ¹⁵N of the three food webs, dietary origins can be traced in the predatory beetle P. japonica derived from wheat, cotton, and maize crops.     

一株产木聚糖酶菌株的筛选鉴定及产酶条件初步优化

本研究从造纸厂的污泥及污水筛选获得了一株产木聚糖酶能力较强的菌株,经鉴定该菌株为异硫链霉菌(Streptomyces althioticus),专利保藏号为CCTCC M 2014110。通过单因素试验初步优化了碳源、氮源、温度、初始p H、发酵周期、摇床转速、接种量及装液量。试验结果显示该菌株最佳产酶条件为:以为玉米芯为碳源,用量30 g/L;以大豆粉与硝酸钾复合物为氮源,用量分别为20g/L和10 g/L;发酵周期120 h,发酵温度40℃,接种量6%,初始p H 5.0,装液量50 m L/250 m L三角瓶,摇床转速为160 r/min。     

Additive and synergistic effects of Bacillus spp. isolates and essential oils on the control of phytopathogenic and saprophytic fungi from medicinal plants and marigold seeds

Contamination of plants and seeds with microorganisms is one of the main problems in the production and distribution of various agricultural products, as well as raw herbal material for the preparation of herbal remedies. In targeting microbial contamination, among other bacteria, Bacillus species showed a significant capacity for biocontrol. The antifungal activity of 14 isolates of Bacillus spp. against 15 fungal isolates from medicinal plants was examined utilizing a dual plate assay. The strongest and broadest antagonistic activity against all fungi tested was exhibited by isolates SS-12.6 and SS-13.1 (from a 43% to 74% reduction in fungal growth), while isolates SS-39.1 and SS-39.3 were effective against the fewest fungus species and also had the weakest antifungal activity. The effect of a crude lipopeptide extract (CLE) of Bacillus sp. SS-12.6 was similar to that achieved by a dual culture with isolate SS-12.6, confirming that the antagonism was the result of the antifungal activities of lipopeptides. In addition, essential oils of thyme (0.55 mg/mL) and savory (0.32 mg/mL) in various combinations with the CLE of SS-12.6 were tested for antifungal activity, and additive and synergistic effects for some of the fungi were obtained. When testing the effect of CLE, oils (0.40 mg/mL for thyme oil and 0.21 mg/mL for savory oil) and combinations in situ on marigold seeds, a reduction of total fungal infection without an adverse effect on germination was accomplished by 6-h treatments with CLE of SS-12.6 (85% reduction of fungal infection and 63% germination), supernatant from liquid culture of SS-12.6 (more than 90% reduction of fungal infection with 69% seed germination) and combinations of CLE and savory oil (77% reduction of fungal infection and 62% seed germination) and CLE with thyme and savory oils (about 75% reduction of fungal infection with 69% seed germination).     

Summer irrigation,grazing and seed addition differentially influence community composition in an invaded serpentine grassland

Community assembly theory predicts that resource availability, biotic interactions, and dispersal dynamics will determine community composition. Recent work has demonstrated that manipulating these processes or “filters” to exclude exotic species may assist in restoring invaded plant communities. In this study, we began by manipulating an abiotic filter, summer water availability, on the theory that irrigation prior to the growing season could trigger the germination of exotic species during unfavorable environmental conditions. First, we performed a greenhouse experiment to assess the germination traits of 23 native and exotic species at low (16°C, spring) and high (30°C, summer) temperatures. At summer temperatures, we found high emergence of many exotic and native grasses and low emergence of native forbs suggesting that summer irrigation may help deplete the exotic seed bank. In a second experiment, we established field plots to test the efficacy of summer irrigation and simultaneously manipulated a biotic and a dispersal filter, subjecting some plots to grazing and/or native seed addition. Summer irrigation and seed addition had no effect on percent cover or species richness while grazing reduced native cover but increased native species richness and soil nitrogen content. Our data suggest that manipulating grazing (a biotic filter) may be more effective than altering abiotic or dispersal filters when restoring invaded serpentine grassland. However, summer irrigation may also be effective, if applied at lower temperatures or for longer periods.