转基因甘蔗BtG-2的T-DNA侧翼序列分析及其转化事件特异性检测

转基因甘蔗BtG-2是利用农杆菌介导法把Cry1Ac-2A-gna融合抗虫基因导入‘新台糖22号’的转基因甘蔗株系,具有良好的抗虫特性和农艺性状。为了明确转基因甘蔗BtG-2的分子特征及其检测方法,推进其生物安全性评价工作,以BtG-2的T2代为研究材料,利用Southern杂交检测外源基因在转基因甘蔗基因组内的拷贝数;利用染色体步移技术分离外源基因在甘蔗基因组中插入位点的侧翼序列,并建立了该转化体高效灵敏的特异性PCR检测方法。结果表明：Southern杂交检测证明外源T-DNA以单拷贝方式插入BtG-2株系;经过3次的热不对称巢式PCR扩增,获得外源基因T-DNA左边侧翼序列984 bp和右边侧翼序列705 bp;以这2个序列和相应的T-DNA的左右端序列分别设计3对检测引物对,建立了BtG-2株系的转化事件特异性PCR检测方法,扩增效率最高的引物对LS011/LA451和RS160/RA588分别扩增到440 bp和428 bp的特异片段。其中T-DNA左侧设计的LS011/LA451检测引物对扩增的灵敏度高、特异性强,能够在甘蔗BtG-2基因组DNA相对含量为0.1%的模板中检测出转基因目的成分,相当于9个单倍体基因组拷贝数。本研究完成了转基因株系BtG-2的分子特征及其转化事件特异性检测,为该转基因甘蔗及其衍生产品的检测和身份识别提供技术依据。     

茶树叶部害虫——茶橙瘿螨

茶橙瘿螨是茶树上一种常见害螨,以成螨、幼螨和若螨刺吸茶树汁液为害茶树,减弱树势,对茶叶产量和品质产生影响。文章介绍了茶橙瘿螨的分布与为害、形态特征、发生规律和防治措施,以期为生产上该害虫的正确识别和有效防治提供参考。     

岩溶流域典型农业区水体氢氧同位素的空间异质性及形成机制

分别于平水期和枯水期采集了花溪河流域典型农业区地表水和地下水样品。利用氢氧同位素示踪技术,结合土地利用类型对研究区不同水体的补给来源、季节变化及主要影响过程进行了分析,并对不同水体氢氧同位素值进行了空间插值分析,同时对其形成机制进行了分析,阐明了不同土地利用类型影响下的主要水文过程。结果表明:(1)研究区不同水体的主要补给来源为当地大气降水,月亮湖水库受蒸发作用影响明显,地表水和地下水的δD和δ~(18)O整体上呈现平水期高于枯水期的特征。(2)地下水的δD和δ~(18)O在枯水期与平水期均呈现明显的空间分异性特征,西部水田/水库集中区富集,东部旱地集中区贫化,土地利用对研究区环境水文过程影响明显。该研究结果有助于了解不同土地利用方式下地表水对地下水的影响,为流域管理提供科学依据。     

Cuticular protein gene LmACP8 is involved in wing morphogenesis in the migratory locust,Locusta migratoria

Cuticular proteins(CPs) are major components of the insect cuticle-associated organs such as integument and wings, although the importance of CPs for wing development and function in hemimetabolous insects remains understudied. In the present study, a wing cuticular protein LmACP8 was identified from Locusta migratoria, which belongs to the RR-2 subfamily of cuticular protein RR consensus(CPR) chitin-binding proteins. LmACP8 was mainly expressed in the wing pads and showed high expression levels before ecdysis of third-, fourth-, and fifth-instar nymphs, with its encoded protein located in the procuticle of wing pads and adult wings. Depletion of LmACP8 by RNA interference markedly reduced the amount of its protein, which consequently caused abnormal wing morphogenesis in the transition from nymph to adult of L. migratoria. We further demonstrated that the abnormal morphogenesis was caused by severe damage of the endocuticle in the wings. LmACP8 was suppressed by 20-hydroxyecdysone(20 E) in vivo, however, its expression was significantly up-regulated after knocking down the hormone receptor gene LmHR39. Thus, the LmACP8 that is negatively regulated by the LmHR39-mediated 20 E signaling pathway is involved in wing development during the nymph to adult transition.     

不同杀青方式龙井茶自动化连续生产线加工工艺探讨

文章剖析了不同杀青方式龙井茶自动化连续生产线的设备配置和工艺流程,对生产线的龙井茶品质进行感官评价,为龙井茶生产线推广应用提供借鉴。     

氨唑草酮的水解、挥发及其在水-沉积物系统中的降解特性

为评价氨唑草酮的环境安全性,参照国家标准GB/T 31270-2014的要求,采用室内模拟法研究了氨唑草酮在不同温度和不同pH值缓冲溶液中的水解特性、在不同环境介质中的挥发特性,以及在2种水-沉积物系统中的降解特性。结果表明:氨唑草酮在25 ℃时,在pH值为4或7的缓冲液中水解半衰期均长于365 d,在pH值为9的缓冲液中水解半衰期为90.0 d,属于难水解至中等水解农药。在20~25 ℃、气体流速500 mL·min^-1的条件下,氨唑草酮在空气、水和土壤中的挥发率均小于1%,属于难挥发农药。氨唑草酮在湖泊(杭州西湖)水-沉积物系统和河流(京杭大运河)水-沉积物系统中的降解符合一级动力学方程,好氧降解半衰期分别为408 d和630 d,厌氧降解半衰期分别为248 d和990 d,在水-沉积物系统中属于难降解农药。     

烤烟控水节肥育苗措施对水肥利用和烟苗素质的影响

为探明有利于减少水肥投入和降低养分残留的烤烟育苗方法,在烤烟育苗环节以常规水肥管理(CM)为对照,设置了控水节肥模式一(OM1)和控水节肥模式二(OM2)两种控水节肥育苗措施,考察了两个试验点不同控水节肥措施下的水肥利用、肥料残留和烟苗生长状况.结果表明,与常规对照(CM)相比,控水节肥(OM)处理下烟株出苗率、茎高、全株生物量及水分利用效率均无显著差异(P>0.05),试验点一和试验点二氮、磷、钾养分残留量分别平均减少52.7％和58.9％、47.8％和60.2％、45.1％和61.5％;与控水节肥模式一(OM1)相比,控水节肥模式二(OM2)耗水量和水分利用效率无显著差异(P>0.05),试验点一和试验点二氮、磷、钾肥残留量分别减少48.6％和28.1％,42.9％和37.7％,36.3％和41.1％,烟苗茎围分别增加了1.8％和27.0％.控水节肥模式二(控制维持水层深度8 cm和初始氮素浓度50 mg/kg)能够在保障烟苗移栽素质的前提下,减少育苗氮肥投入47.7％,降低苗池水体氮、磷、钾养分残留量,是一种值得推荐的绿色高效育苗水肥管理措施.     

Screening and Verification of Low Nitrogen Tolerant and Nitrogen Sensitive Cotton Germplasm

[Objective] The aim of this study is to evaluate nitrogen efficient cotton germplasms and improve nitrogen use efficiency. [Method] Eighty cotton germplasms were selected and evaluated in the hydroponic experiment under low (0.25 mmol·L^－1) and high (5 mmol·L^－1) nitrogen concentration. Different traits for screening were identified and nitrogen use efficiency types were classified. Field experiments were also performed for comparison and confirmation of the identified germplasms. [Result] The results showed that there were significant differences in the total plant dry matter, shoot nitrogen accumulation and nitrogen absorption efficiency in cotton germplasms at the two nitrogen levels. Based on coefficient of variation, principal component analysis and correlation, six traits including total plant dry matter, shoot dry matter, root dry matter, total nitrogen accumulation, shoot nitrogen accumulation and nitrogen absorption efficiency were used as screening indicators. According to the Heatmap clustering analysis and the nitrogen efficiency comprehensive index, two germplasms (Lu05R59 and CCRI 69) were identified as low nitrogen tolerant and nitrogen efficient, and two germplasms (Coker 201 and Xinluzhong 30) as low nitrogen sensitive and nitrogen inefficient. The results of field experiment were consistent with the results of the hydroponic culture at the seedling stage. [Conclusion] It was finally determined that Lu05R59 and CCRI 69 were the low nitrogen tolerant and nitrogen efficient germplasms, and Coker 201 and Xinluzhong 30 were low nitrogen sensitive and nitrogen inefficient germplasms. The results of these studies provide the possibility for screening and rapid identification of nitrogen use efficiency in cotton at the seedling stage, and provide the ideal materials and theoretical basis for further study of cotton nitrogen efficient.     

Regulatory role of l-proline in fetal pig growth and intestinal epithelial cell proliferation

l-proline (Pro) is a precursor of ornithine, which is converted into polyamines via ornithine decarboxylase (ODC). Polyamines plays a key role in the proliferation of intestinal epithelial cells. The study investigated the effect of Pro on polyamine metabolism and cell proliferation on porcine enterocytes in vivo and in vitro. Twenty-four Huanjiang mini-pigs were randomly assigned into 1 of 3 groups and fed a basal diet that contained 0.77% alanine (Ala, iso-nitrogenous control), 1% Pro or 1% Pro + 0.0167% α-difluoromethylornithine (DFMO) from d 15 to 70 of gestation. The fetal body weight and number of fetuses per litter were determined, and the small and large intestines were obtained on d 70 ± 1.78 of gestation. The in vitro study was performed in intestinal porcine epithelial (IPEC-J2) cells cultured in Dulbecco''s modified Eagle medium-high glucose (DMEM-H) containing 0 μmol/L Pro, 400 μmol/L Pro, or 400 μmol/L Pro + 10 mmol/L DFMO for 4 d. The results showed that maternal dietary supplementation with 1% Pro increased fetal weight; the protein and DNA concentrations of the fetal small intestine; and mRNA levels for potassium voltage-gated channel, shaker-related subfamily, member 1 (Kv1.1) in the fetal small and large intestines (P < 0.05). Supplementing Pro to either gilts or IPEC-J2 cells increased ODC protein abundances and polyamine concentrations in the fetal intestines and IPEC-J2 cells (P < 0.05). In comparison with the Pro group, the combined administration of Pro and DFMO reduced the expression of ODC protein and spermine concentration in the fetal intestine, as well as the concentrations of putrescine, spermidine and spermine in IPEC-J2 cells (P < 0.05). Meanwhile, the percentage of cells in the S-phase and the mRNA levels of proto-oncogenes c-fos and c-myc were increased in response to Pro supplementation, whereas depletion of cellular polyamines with DFMO increased tumor protein p53 (p53) mRNA levels (P < 0.05). Taken together, dietary supplementation with Pro improved fetal pig growth and intestinal epithelial cell proliferation via enhancing polyamine synthesis.     

体外产气法评价全株藜麦与全株玉米混合青贮饲料饲用价值

为提高全株藜麦的饲料化利用率，寻找其与全株玉米的最佳青贮比，试验共设全株藜麦与全株玉米比为100：0，90：10，80：20等11个组合进行青贮发酵，发酵60 d后测定各组青贮料的体外产气发酵指标，使用灰色关联度分析筛选出最佳青贮比例。结果表明，体外发酵至36 h时，各处理组GP趋于稳定，体外发酵减弱接近停止；全株藜麦与全株玉米比例为90：10时，该青贮饲料组合的粗蛋白（CP）和粗脂肪（EE）含量显著高于其他各组（P<0.05），乙酸（AA）、总挥发性脂肪酸（TVFA）含量以及氨态氮（NH₃-N）浓度显著高于其他各处理（P<0.05）。分析表明，全株藜麦与全株玉米比例为90：10时，混合青贮饲料的营养品质和产气发酵特性综合表现最好，可作为优质混合青贮饲料在家畜养殖中推广使用。