木薯(Manihot esculenta)AKT1基因的克隆及生物信息学分析

植物细胞中的K^+通道蛋白AKT1(Arabidopsis K^+transpoter 1)主要负责吸收K^+。本研究基于木薯基因组数据库信息,利用PCR技术从木薯中克隆一个MeAKT1基因,该基因全长2634 bp,编码877个氨基酸,生物信息学分析表明MeAKT1含有10个外显子和9个内含子,编码的氨基酸分子量为99.02 kD,等电点为8.43,属于稳定蛋白。MeAKT1包含有5个跨膜区域,N端含有1个Ion_trans结构域,C端有1个KHA结构域,其二级结构以α-螺旋和无规则卷曲为主,进化树分析发现MeAKT1与蓖麻RcAKT1的亲缘关系最近。通过对MeAKT1蛋白的生物信息学分析有助于下一步深入研究MeAKT1在木薯耐贫瘠过程中的功能。     

大丽轮枝孢激活蛋白对小麦旗叶面积及其光合特性和籽粒产量的影响

为明确大丽轮枝孢激活蛋白（Verticillium Dahliae Asp-f2 Like，简称VDAL）对冬小麦旗叶面积、花后光合特性及籽粒产量的影响，于2017-2018和2018-2019年两个小麦生长季，以强筋冬小麦品种藁优2018为材料，在河北藁城设置大丽轮枝孢激活蛋白种子包衣，并在大田条件下设置春三叶（倒四叶）露尖（S3）、春四叶（倒三叶）露尖（S4）、春五叶（倒二叶）露尖（S5）、春六叶（旗叶）露尖（S6）以及孕穗期（SB）叶面喷施VDAL和全生育期不喷施VDAL（S0）6个处理，以不包衣且不喷施为对照（CK），分析了VDAL对小麦旗叶面积、花后光合特性和产量的影响。结果表明，S4和S5处理显著（P＜0.05）增加了旗叶的长或宽，从而增大了叶面积。S4、S5和S6处理可以维持旗叶花后较高水平的净光合速率。各喷施处理的籽粒产量均有不同程度的提高，与CK相比，两年间S4处理的籽粒产量均显著（P＜0.05）提高，2018-2019年，S5处理的穗粒数和籽粒产量也显著（P＜0.05）提高。综上，采用VDAL种子包衣，并于春生4~5叶龄喷施VDAL，可以增加旗叶面积和旗叶花后净光合速率，提高穗粒数和千粒重，从而达到增加产量的目的。     

草坪草屑的加工与利用

我国每年修剪草坪产生大量的草坪草屑,对草坪草屑的加工利用,可以对我国能源、饲料、肥料及蛋白资源的短缺起到一定的缓解作用,因此如何对草坪草屑进行合理利用已成为一项重要的研究课题。本文综述了草坪草屑能源化、饲料化、肥料化和深加工4种利用方式,详细介绍了草坪草屑在发酵生产能源、饲喂家畜、堆肥还坪以及加工叶蛋白等方面的研究进展,指出禾本科草坪草屑和豆科草坪草屑应采用不同的处理方式,对各加工方式的投入和产出也应进行进一步的量化核算,同时总结并展望了草坪草屑利用方式的发展前景。     

稻瘟病菌F-box蛋白MoFbr7生物学功能分析

F-box蛋白在泛素-蛋白酶体途径(ubiquitin-proteasome pathway,UPP)中参与调控胞内蛋白降解、受体识别、信号传导等生物学过程。本研究从稻瘟病菌中克隆了F-box基因MoFbr7,序列分析表明,该基因编码产物具有1个F-box结构域(N-端)和8个连续的WD40重复序列(C-端),在丝状真菌中高度保守。利用基因敲除方法,获得4个MoFbr7基因敲除突变体,同时构建了回补菌株。表型分析结果显示,MoFbr7基因缺失突变体在产孢量、附着胞形态、原生质体释放、致病性等方面均无异常。突变体在MM、RDC培养基上生长速率下降;对细胞壁胁迫因子CFW(Calcofluor white)、刚果红敏感。以上结果表明MoFbr7参与稻瘟病菌的营养生长与细胞壁完整性,为进一步揭示其生物学功能奠定基础。     

Effects of Dietary Protein and Lipid Levels with Different Protein‐to‐Energy Ratios on Growth,Feed Utilization,and Plasma Biochemical Parameters of Dianchi Golden‐line Barbell,Sinocyclocheilus grahami

An 8‐wk feeding trial was conducted to evaluate the effects of varying dietary protein and lipid levels with different protein‐to‐energy (P/E) ratios on growth, feed utilization, and plasma biochemical parameters of Sinocyclocheilus graham. Nine experimental diets were formulated to contain three protein levels (35, 40, and 45%), and each with three lipid levels (4, 8, and 12%), to produce a range of P/E ratios from 71.31 to 98.64 mg protein/kcal. Each diet was randomly assigned to triplicate groups of 25 fish. Fish grew best when fed the diet containing 45% protein and 12% lipid. However, fish fed the diet with 40% protein and 8% lipid had similar weight gain and specific growth rate as those fed the above diet but showed relatively better feed conversion ratio, protein efficiency ratio, and protein and energy retentions. Additionally, plasma total amino acids and blood urea nitrogen contents and γ‐glutamyltransferase, aspartate aminotransferase, and alanine aminotransferase activities in fish fed the diet containing 40% protein and 8% lipid were relatively low among the dietary treatments. These results may indicate that a diet containing 40% protein and 8% lipid with P/E ratio of 84.05 mg protein/kcal would be suitable for the optimal growth, feed utilization, and health of S. graham.     

对草地贪夜蛾高毒力的苏云金芽胞杆菌菌株筛选与杀虫活性研究

草地贪夜蛾是一种世界性重大农业害虫，给玉米等多种主要粮食和经济作物造成严重危害。为实现对草地贪夜蛾的高效、绿色、持续防控，亟需筛选高毒力苏云金芽胞杆菌菌株资源。本研究以前期实验室储备的对斜纹夜蛾、小地老虎具有杀虫活性的363株野生菌株库为候选菌株，基于菌株杀虫基因差异及进化关系，去除冗余菌株后获得172株候选菌株。通过培养获取这些菌株胞晶混合物，进行杀虫蛋白定量后，测定杀虫活性，获得27株对草地贪夜蛾初孵幼虫具有较高杀虫活性的菌株，其中菌株B14-D2的杀虫活性最高，LC₅₀为0.155 μg/g。克隆了该菌株中的cry1Ea3基因并在HD73^-无晶体突变株中表达，Cry1Ea3蛋白对草地贪夜蛾初孵幼虫LC₅₀为1.789 μg/g。本研究为新的Bt产品和杀虫基因的开发利用提供了丰富资源。     

猪链球菌蛋白表面展示系统的构建及展示蛋白的初步观察

为构建猪链球菌(Streptococcus suis)蛋白表面展示系统,本研究通过序列分析,确定猪链球菌的LPxTG蛋白及其信号肽(SP)和胞壁锚定基序(CWA),通过PCR扩增Peno-SP、GFP、CWA的DNA片段并融合,构建强启动子Peno控制表达编码SP-GFP-CWA融合蛋白的DNA片段,将该重组DNA片段连接pSET2载体,获得蛋白表面展示质粒,转化猪链球菌,构建得到以GFP为报告蛋白的猪链球菌蛋白表面展示系统。结果显示,利用猪链球菌的10个LPxTG蛋白及其SP和CWA序列,构建了10个含有Peno-SP-GFP-CWA融合片段的重组pSET2表面蛋白展示质粒pSsPSD1至pSsPSD10,分别转化猪链球菌05ZYH33,PCR鉴定显示其中7个转化猪链球菌。采用western blot初步检测其展示蛋白,结果显示,7个转化阳性菌株均能有效表达GFP蛋白,以成熟GFP条带为指标,均表现出了一定的外源GFP表面展示水平,分别命名为SsPSD1、SsPSD2、SsPSD4、SsPSD7-SsPSD10,其中SsPSD1、SsPSD4、SsPSD8和SsPSD9表面展示水平相对较好,在猪链球菌表面展示外源蛋白方面具有很好的潜力。本研究首次尝试建立猪链球菌蛋白表面展示系统,为猪链球菌表面递呈外源蛋白或抗原提供了新的策略。     

Growing Degree Day: Noninvasive Remotely Sensed Method to Monitor Diet Crude Protein in Free-Ranging Cattle

The daily nutritional balance of free-ranging cattle is the net result of intake from available forage biomass and nutritive value weighed against the nutritional requirements of the animal. Plant phenology influences nutritive value. Plant phenology is dictated by time of year and an accumulation of photosynthetically active days. Growing degree day (GDD) is a concept that quantifies this relationship and has been used to predict nutritive value in perennial range grasses. GDD could be substituted for chemical analysis to inform grazing animal nutritional monitoring efforts. We hypothesized that in C4 grass-dominated rangelands, a cumulative GDD calculation would correlate with diet crude protein (CP) predictions obtained by fecal near infrared spectroscopy (FNIRS) from free-ranging cattle. Therefore, the objectives of our research were to evaluate the effectiveness of GDD to predict FNIRS-derived determinations of grazing cattle diet CP in 1) two groups of three individual animals grazing a small native pasture and 2) large commercial-scale herds grazing expansive rangelands. For the first objective, cumulative GDD and FNIRS-predicted diet CP were strongly correlated (r² = 0.76; P < 0.01). Relationships between cumulative GDD and FNIRS-predicted diet CP for the second objective varied considerably among ranches, ranging from a low r² of 0.05 (P = 0.871) to a high r² of 0.78 (P < 0.049). Similar values for individual ranch/year combinations were stronger; ranging from a minimum r² of 0.44 (P = 0.556) to a maximum of 0.95 (P = 0.051). The aggregate relationship between GDD and FNIRS-predicted CP for all ranch/year combinations was highly significant (r² = 0.37; P < 0.001), but the standard error was 1.86% CP. The noninvasive remotely sensed grazing animal nutritional monitoring method described here was accurate enough to inform tactical rangeland diet quality assessments but was not accurate enough to inform operational-scale grazing management decisions.