甘蔗热带种金属硫蛋白家族基因的克隆及响应重金属胁迫的表达分析

金属硫蛋白是一类富含巯基的低分子量蛋白,在植物的重金属解毒及细胞氧化还原调控等方面起重要的作用。本研究以甘蔗热带种Badila组培苗为材料,分别测定了其在CdCl2、ZnSO4和CuCl2水溶液培养条件下地上部和地下部的重金属含量,结果显示其对上述3种重金属有较强的耐受与富集能力。继而克隆了ScMT1(登录号为KJ504373)、ScMT2-1-5(登录号为MH191346)和ScMT3(登录号为KJ5043704)3个金属硫蛋白家族基因,它们分别属于植物MT亚家族中的MT1、MT2和MT3型基因。ScMT1含有1个内含子和2个外显子,开放阅读框(Open Reading Frame,ORF)长228 bp,编码75个氨基酸;ScMT2-1-5含有2个内含子和3个外显子,ORF长246 bp,编码81个氨基酸;ScMT3含有1个内含子和2个外显子,ORF长198 bp,编码65个氨基酸。RT-qPCR显示,Cd2+胁迫下,在甘蔗地上部和地下部,ScMT2-1-5均连续显著上调表达,而ScMT1的上调应答出现延迟。ScMT3在地上部的上调应答出现延迟,在地下部呈“扬-抑”趋势,提示甘蔗响应Cd^2+胁迫过程中ScMT2-1-5起更积极的作用,ScMT1参与胁迫后期的分子响应,而ScMT3不起主导作用。Cu^2+胁迫下,地上部ScMT1连续显著上调表达,ScMT2-1-5和ScMT3呈总体上调的表达趋势;地下部,ScMT1和ScMT2-1-5的上调表答均出现延迟,仅在胁迫后期显著上调表达,而ScMT3仅在胁迫前期显著上调表达。该结果提示了ScMT1、ScMT2-1-5和ScMT3在Cu2+胁迫响应过程中的协作关系,三者共同参与了地上部的胁迫响应,其中ScMT1起更积极的作用;此外三者还先后参与了地下部对Cu^2+胁迫的分子响应。Zn^2+胁迫下,ScMT1和ScMT3分别仅在地上部和地下部显著上调表达;ScMT2-1-5在地上部和地下部均呈“扬-抑”的应答趋势;提示了在甘蔗响应Cd^2+胁迫应答过程中ScMT1和ScMT3分别在地上部和地下部起主要作用,ScMT2-1-5参与了胁迫前期的分子响应。ScMT1、ScMT2-1-5和ScMT3在甘蔗不同组织中及在重金属(Cd^2+、Zn^2+或Cu^2+)不同累积水平下呈现出相似或互补的应答特性,提示上述甘蔗MT家族不同成员在重金属解毒及细胞氧化还原调控等方面产生了功能分化,且三者在应对过量Cd^2+、Zn^2+或Cu^2+对甘蔗组织造成伤害的过程中存在时空上的协同作用。该研究为深入理解多倍体植物甘蔗中MT家族各成员基因在重金属耐受过程中的协同作用机制奠定了基础。     

茶树花青素形成分子机理的研究进展

花青素是由类黄酮合成途径产生的次生代谢产物,含量高时会使茶树新梢呈现红色或紫色。同时,花青素相比儿茶素等具有更明显的抗氧化、预防肿瘤等药理保健作用。文章就茶树花青素合成途径、转录及转录后调控等方面进行综述,以期更好地为高花青素茶树的育种研究提供理论依据。     

Exogenous application of allelopathic water extracts helps improving tolerance against terminal heat and drought stresses in bread wheat (Triticum aestivum L. Em. Thell.)

The allelopathic water extracts (AWEs) may help improve the tolerance of crop plants against abiotic stresses owing to the presence of the secondary metabolites (i.e., allelochemicals). We conducted four independent experiments to evaluate the influence of exogenous application of AWEs (applied through seed priming or foliage spray) in improving the terminal heat and drought tolerance in bread wheat. In all the experiments, two wheat cultivars, viz. Mairaj‐2008 (drought and heat tolerant) and Faisalabad‐2008 (drought and heat sensitive), were raised in pots. Both wheat cultivars were raised under ambient conditions in the wire house till leaf boot stage (booting) by maintaining the pots at 75% water‐holding capacity (WHC). Then, managed drought and heat stresses were imposed by maintaining the pots at 35% WHC, or shifting the pots inside the glass canopies (at 75% WHC), at booting, anthesis and the grain filling stages. Drought stress reduced the grain yield of wheat by 39%–49%. Foliar application of AWEs improved the grain yield of wheat by 26%–31%, while seed priming with AWEs improved the grain yield by 18%–26%, respectively, than drought stress. Terminal heat stress reduced the grain yield of wheat by 38%. Seed priming with AWEs improved the grain yield by 21%–27%; while foliar application of AWEs improved the grain yield by 25%–29% than the heat stress treatment. In conclusion, the exogenous application of AWEs improved the stay green, accumulation of proline, soluble phenolics and glycine betaine, which helped to stabilize the biological membranes and improved the tolerance against terminal drought and heat stresses.     

Mapping the spatial distribution of shade provision of street trees in Boston using Google Street View panoramas

Street trees provide shade and increase human thermal comfort during hot summer. In this study, we investigated the spatial distribution of shade provision of street trees in Boston, Massachusetts. The sky view factor (SVF), which influences the solar radiation to the ground and affects human thermal comfort, was used to indicate the contribution of street trees on shade provision. Google Street View (GSV) panoramas were used to calculate the photographic method based SVF (SVF_P), with the consideration of all kinds of obstructions within street canyons. A building height model was used to calculate the simulation based SVF (SVFs), with consideration of obstruction of building blocks only. Considering the fact that street trees and building blocks are the two major obstructions of radiation within street canyons, therefore, the difference between the two SVF estimation results can be considered as the shade provision of street trees. The results show that street trees help to decrease the SVF by 24.61% in Boston, Massachusetts. The shading level varies spatially in the study area. Generally, the southwestern area has much higher shading level than the north and the east. We further explored the shading variation among different socioeconomic groups in the study area. Result shows that Hispanics tend to live in neighborhoods with lower shading level. This study can help to provide a reference for future urban greening projects for global climate change adaption.     

品种间大豆脂肪氧化酶活性差异及β-胡萝卜素漂白性研究

为了解东北地区大豆品种间大豆脂肪氧化酶(LOX)活性差异规律,整理适于东北地区种植的大豆种质资源307份(6份半野生品种,17份地方品种及4份国外品种,其余为栽培品种),统一种植,秋季收获后,测定各品种成熟籽粒脂肪氧化酶总活性。分析品种间脂肪氧化酶总活性分布规律、大豆脂肪氧化酶总活性与β-胡萝卜素漂白性之间关系。结果表明,307份大豆品种间脂肪氧化酶总活性差异极显著,平均值为21.68 U·mL~(-1),最小值为2.51 U·mL~(-1),最大值为42.38 U·mL~(-1)。根据脂肪氧化酶总活性差异,通过聚类分析将307份大豆品种分为3类。低活性品种占总量43.65%,脂肪氧化酶总活性平均值为15.25 U·mL~(-1);中等活性品种占总量43.97%,脂肪氧化酶活性平均值为24.64 U·mL~(-1);高活性品种占总量12.38%,脂肪氧化酶总活性平均值为33.87 U·mL~(-1)。脂肪氧化酶总活性与油分含量呈正相关,与蛋白含量呈负相关,与漂白性呈正相关。     

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。     

马疱疹病毒1型主要毒力基因分析及TK基因缺失载体的构建

为了解新疆马疱疹病毒1型（EHV-1）主要毒力基因遗传进化情况并构建TK基因缺失株，本研究以EHV-1 XJ2015株DNA为模板，对其主要毒力基因TK、gI和gE全长进行克隆、测序及生物信息学分析，并扩增TK基因左右重组臂TKL和TKR，构建质粒pUC-TKLR，将扩增后的增强绿色荧光蛋白（EGFP，含有CMV+polyA）插入pUC-TKLR质粒，构建TK基因缺失打靶质粒。TK、gI和gE基因同源性分析结果显示，XJ2015株与国外EHV-1分离株TK、gI和gE基因同源性均较高，分别为99.8%~100.0%、99.6%~100.0%和99.9%~100.0%；与EHV-3分离株同源性均最低，分别为72.9%、59.4%和62.1%；遗传进化分析显示，3个基因均与国外EHV-1同属于一个遗传进化分支，与EHV-9和EHV-4进化关系较近，但与EHV-3进化关系较远，表明XJ2015毒株与国外EHV-1毒株TK、gI、gE基因核苷酸上差异不明显，没有明显的地域性特征，功能基因保守且进化缓慢，同源基因功能相同或相近；经PCR扩增、酶切、测序及转染鉴定，本试验成功构建了用于TK基因缺失的打靶质粒pUC-TKLR-EGFP。通过对EHV-1主要毒力基因的分析及TK基因缺失打靶载体的构建，为新疆地区马鼻肺炎流行病学调查分析、TK基因缺失株的构建提供理论依据。     

A comparison of the effects of carbon dioxide and medical air for abdominal insufflation on respiratory parameters in xylazine-sedated sheep undergoing laparoscopic artificial insemination

AIMS: To determine if abdominal insufflation with medical air will improve oxygenation and ventilation parameters when compared to insufflation with CO₂ in xylazine-sedated sheep undergoing laparoscopic artificial insemination (AI).

METHODS: Forty-seven sheep underwent oestrus synchronisation and were fasted for 24 hours prior to laparoscopic AI. Each animal was randomised to receive either CO₂ or medical air for abdominal insufflation. An auricular arterial catheter was placed and utilised for serial blood sampling. Respiratory rates (RR) and arterial blood samples were collected at baseline, after xylazine (0.1?mg/kg I/V) sedation, 2 minutes after Trendelenburg positioning, 5 minutes after abdominal insufflation, and 10 minutes after being returned to a standing position. Blood samples were collected in heparinised syringes, stored on ice, and analysed for arterial pH, partial pressure of arterial O₂ (PaO₂), and CO₂ (PaCO₂). The number of ewes conceiving to AI was also determined.

RESULTS: Repeated measures ANOVA demonstrated temporal effects on RR, PaO₂, PaCO₂ and arterial pH during the laparoscopic AI procedure (p<0.001), but no difference between insufflation groups (p>0.01). No sheep experienced hypercapnia (PaCO₂>50?mmHg) or acidaemia (pH<7.35). Hypoxaemia (PaO₂<70?mmHg) was diagnosed during the procedure in 14/22 (64%) ewes in the CO₂ group compared with 8/23 (35%) ewes in the medical air group (p=0.053). Overall, 15/20 (75%) ewes in the CO₂ group conceived to AI compared with 16/22 (72.7%) in the medical air group (p=0.867).

CONCLUSIONS AND CLINICAL RELEVANCE: There were no statistical or clinical differences in RR, PaO₂, PaCO₂, pH, or conception to AI when comparing the effects of CO₂ and medical air as abdominal insufflation gases. None of the sheep experienced hypercapnia or acidaemic, yet 42% (19/45) of sheep developed clinical hypoxaemia, with a higher percentage of ewes in the CO₂ group developing hypoxaemia than in the medical air group. Based on the overall analysis, medical air could be utilised as a comparable alternative for abdominal insufflation during laparoscopic AI procedures.