Effects of different NS genes of avian influenza viruses and amino acid changes on pathogenicity of recombinant A/Puerto Rico/8/34 viruses

To examine the effects of the NS1 and NEP genes of avian influenza viruses (AIVs) on pathogenicity in mice, we generated recombinant PR8 viruses containing 3 different NS genes of AIVs. In contrast to the reverse genetics-generated PR8 (rPR8) strain and other recombinant viruses, the recombinant virus rPR8-NS(0028), which contained the NS gene of A/chicken/KBNP-0028/2000 (H9N2) (0028), was non-pathogenic to mice. The novel single mutations of 0028 NS1 to corresponding amino acid of PR8 NS1, G139D and S151T increased the pathogenicity of rPR8-NS(0028). The replacement of the PL motifs (EPEV or RSEV) of pathogenic recombinant viruses with that of 0028 (GSEV) did not reduce the pathogenicity of the viruses. However, a recombinant virus with an EPEV-grafted 0028 NS gene was more pathogenic than rPR8-NS(0028) but less than rPR8. The lower pathogenicity of rPR8-NS(0028) might be associated with the lower virus titer and IFN-β level in the lungs of infected mice, and be attributed to G139, S151 and GSEV-PL motif of NS1 gene of 0028. In conclusion we defined new amino acid residues of NS1 related to mice pathogenicity and the presence of pathogenic NS genes among low pathogenic AIVs may encourage continuous monitoring of their mammalian pathogenicity.     

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。     

3D-MOTIF Method and Its Application to Microscale Topography

The 2D motif method is a surface characterization method promising to separate roughness and waviness from a profile and is adopted by ISO,so is meaningful to extend this method to 3D.Vincent's watershed algorithm is employed for the generation of 3D-motif.A smallest area selecting criteria is proposed for the use of clearing the small motifs which are concerned as noise.As there are no manipulating factors as 2D-method,a multi-scale analysis is employed based on area and depth criteria,the use of depth criteria is to prevent the combination of two adjacent motifs if there is a significant peak on the border of them.Finally the surface of C(100) are analyzed by the presented method,the texture of this surface has been characterized.     

Complexity of indica-japonica varietal differentiation in Bangladesh rice landraces revealed by microsatellite markers

To understand the genetic diversity and indica-japonica differentiation in Bangladesh rice varieties, a total of 151 accessions of rice varieties mostly Bangladesh traditional varieties including Aus, Boro, broadcast Aman, transplant Aman and Rayada varietal groups were genotyped using 47 rice nuclear SSRs. As a result, three distinct groups were detected by cluster analysis, corresponding to indica, Aus and japonica rice. Among deepwater rice varieties analyzed some having particular morphological features that mainly corresponded to the japonica varietal group. Some small seeded and aromatic varieties from Bangladesh also corresponded to the japonica varietal group. This research for the first time establishes that the japonica varietal group is a prominent component of traditional varieties in Bangladesh, particularly in deepwater areas.     

SSR分子标记在苹果研究中的应用

SSR(simple sequence repeat)分子标记是以聚合酶链式反应(PCR)技术为基础的共显性分子标记,也称为微卫星标记(microsatellite markers),其具有多态性高、通用性好、易检测且操作简单等优点,已被较多应用于苹果的品种鉴定、遗传多样性分析以及遗传图谱构建和基因定位等方面。就以上方面对苹果中SSR分子标记的应用进行概述并提出SSR分子标记在未来苹果中的研究重点和方向。     

屡配不孕母牛发情开始后八天内血清促黄体素（LH）含量的变化

本实验采用放射免疫测定技术(RIA)分析了9头屡配不孕母牛和6头正常母牛于发情开始后8天内的血清促黄体素(LH)的含量变化。测定结果表明:屡配不孕母牛的LH排卵峰值低于正常母牛,其峰值分别为2.71±1.00ng/ml和4.70±1.71ng/ml,差异显著(P<0.025)。为了初步验证LH峰值与屡配不孕之间的关系,对20头屡配不孕母牛在发情开始至配种前肌肉注射外源性LH180IU,其配种后60天的受胎率为70％(14/20),而未用LH处理的11头屡配不孕母牛的受胎率为18％(2/11),两者差异极显著(P<0.01)。上述实验结果提示:体内LH排卵峰值降低可能是导致乳牛屡配不孕的重要原因之一。     

绿豆种质资源苗期抗旱性鉴定

鉴定筛选苗期抗旱绿豆种质,对改良绿豆品种抗旱性、促进绿豆产业发展具有重要意义,同时,也为"绿豆抗旱性鉴定评价技术规范"的制定提供方法和信息支撑。本研究以70份绿豆种质为材料,采用苗期反复干旱法,测定幼苗存活率、萎蔫指数、株高、叶片鲜重、叶片干重、叶片含水量、生物量和胁迫指数等指标,分析各指标间的相关性,结果显示第1次旱胁迫后的幼苗存活率与第2次旱胁迫后的幼苗存活率呈极显著正相关;萎蔫指数、株高与幼苗存活率呈极显著负相关,故遴选出第1次旱胁迫幼苗存活率、萎蔫指数和株高为绿豆苗期抗旱性评价的适宜指标。采用隶属函数法,综合分析划级,获得高抗旱种质16份、抗旱种质20份,中抗种质23份、敏感种质8份和极敏感种质3份。以第1次旱胁迫幼苗存活率、萎蔫指数和胁迫株高分别评价绿豆的抗旱性并与综合评价结果相比较,一致率分别为70.0%、58.6%和51.4%。认为第1次旱胁迫幼苗存活率可以作为大规模绿豆种质苗期抗旱筛选的鉴定指标。     

鸡败血支原体pMGA多基因家族及其免疫逸机制的研究进展

pMGA多基因家族主要编码一类黏附素／血凝素蛋白，存在于鸡败血支原体的细胞表面，其主要功能是促进支原体黏附到宿主细胞上。近年来的相关研究发现，编码pMGA的基因数量从32－70不等，主要转录水平上通过（GAA）n 基序的数量改变引发pMGA基因的选择性转录，造成pMGA的抗原发生变异，从而干扰宿主正常免疫功能的发挥，使支原体对宿主产生严重的免疫逃逸。其中，（GAA）n基序已被证实在pMGA基因表达调控中起重要作用，这一三联体重复基序数目的多少直接影响到pMGA基因的ON／OFF不，本文通过对鸡败血支原体pMGA多基因家分子生物学研究进展浅要综述，以提出pMGA基因可能的转录调控机制，这对研究鸡败血支原体的分子致病机理及其免疫机制大有裨益。     

P0 protein of cotton leafroll dwarf virus-atypical isolate is a weak RNA silencing suppressor and the avirulence determinant that breaks the cotton Cbd gene-based resistance

Cotton blue disease (CBD) is the most important disease present in cotton crops in South America and cotton leafroll dwarf virus (CLRDV) is the causal agent. The disease has been controlled by sowing cotton varieties resistant to CLRDV. However, in the 2009/10 growing season, an outbreak due to an atypical CLRDV isolate (CLRDV-at) occurred in northwest Argentina. Although CLRDV and CLRDV-at genomes are very closely related, the symptoms they produce in cotton plants are quite different. P0 is the most divergent protein between the isolates and in CLRDV is a silencing suppressor protein. This work characterized the silencing suppressor activity of the P0 protein encoded by CLRDV-at (P0^CL-at) and evaluated its role in Cbd-resistance break in cotton plants. It was demonstrated that P0^CL-at, despite having a mutation in the consensus of the F-box-like motif, was able to suppress local RNA silencing, but displayed lower activity than P0^CL. P0^CL and P0^CL-at showed no differences in the interaction with Gossypium hirsutum SKP1 orthologue (GSK1) and Nicotiana benthamiana SKP1 and both P0 proteins triggered destabilization of ARGONAUTE1. However, when the ability to enhance PVX symptoms was evaluated, P0^CL-at was shown to be a weaker pathogenicity factor than P0^CL in N. benthamiana. Interestingly, trans-expressed P0^CL-at enabled CLRDV to systemically infect CBD-resistant plants, and a chimeric CLRDV-P0^CL-at infectious clone succeeded in establishing infection in CBD-resistant cotton varieties with symptoms resembling those produced by CLRDV-at. These results strongly suggest that P0^CL-at is the avirulence (Avr) determinant involved in breaking cotton Cbd gene-based resistance.