大花君子兰叶绿体基因组及其特征

采用Illumina MiSeq测序平台对大花君子兰（Clivia miniata）叶片总DNA进行测序，通过组装获得了其叶绿体基因组（cpDNA）全长序列（158 114 bp）。对其cpDNA注释得到135个基因，包含87个蛋白编码基因、40个tRNA基因和8个rRNA基因。采用生物信息学方法对获得的cpDNA进行简单序列重复（SSR）分析和密码子偏好性分析。结果显示：①大花君子兰cpDNA中共有61个SSR位点，其中单核苷酸、二核苷酸、三核苷酸、四核苷酸、五核苷酸和六核苷酸重复数分别为38、9、2、8、3和1个，多数SSR分布在基因间隔区；②大花君子兰cpDNA密码子偏爱以A或U（T）结尾，亮氨酸使用频率最高，半胱氨酸使用频率最低。基于24种植物的cpDNA全长和23种植物的叶绿体ycf2基因序列进行系统发育分析，结果显示大花君子兰与石蒜科植物在同一分支，显示最近的亲缘关系，支持大花君子兰属于石蒜科。基于叶绿体ycf2的系统发育分析结果与基于cpDNA全长的系统发育分析研究结果大部分相同，支持ycf2基因可以代替cpDNA全长用于植物系统发育分析。     

玉米开花期相关的Indeterminate domain(IDD)蛋白家族基因的鉴定

开花期是影响玉米产量的重要因子之一。Indeterminate1 (ID1)编码玉米Indeterminate domain (IDD)家族蛋白,是玉米开花期的重要调控因子。然而,其他玉米IDD蛋白家族基因及其生物学功能有待深入研究。本文利用生物信息学技术在玉米基因组中鉴定并分离了37个IDD家族基因,记作ZmIDD。表达分析发现这些ZmIDD基因在8个玉米组织中显示出多种表达模式。为进一步探讨ZmIDD基因在调控玉米开花期上的作用,检测了37个ZmIDD在172个自交系中的遗传多样性,发现35个ZmIDD基因在自交系间具有多态性,平均每个基因具有37.8个多态性位点。关联分析鉴定到包含ID1在内的7个ZmIDD基因在多个环境下与开花期性状显著关联。对Zm00001d020683基因2 kb的启动子区和600 bp编码区重测序,共鉴定到64个多态性位点。候选基因关联分析鉴定到2个启动子区的插入缺失(In/Del)位点与开花期显著关联,其中2个位点分别插入3 bp和2 bp的单倍型为一种提早开花的基因型。研究结果为玉米开花期相关基因的分离和利用研究提供了候选基因和选择靶点。     

A Lymphoid Organ Specific Anti-Lipopolysaccharide Factor from Litopenaeus vannamei Exhibits Strong Antimicrobial Activities

Different shrimp species are known to possess apparent distinct resistance to different pathogens in aquaculture. However, the molecular mechanism underlying this finding still remains unknown. One kind of important antimicrobial peptides, anti-lipopolysaccharide factors (ALF), exhibit broad-spectrum antimicrobial activities. Here, we reported a newly identified ALF from the shrimp Litopenaeus vannamei and compared the immune function with its counterpart in the shrimp Fenneropenaeus chinensis. The ALF, designated as LvALF8, was specifically expressed in the lymphoid organ of L. vannamei. The expression level of LvALF8 was apparently changed after white spot syndrome virus (WSSV) or Vibrio parahaemolyticus challenges. The synthetic LBD peptide of LvALF8 (LvALF8-LBD) showed strong antibacterial activities against most tested Gram-negative and Gram-positive bacteria. LvALF8-LBD could also inhibit the in vivo propagation of WSSV similar as FcALF8-LBD, the LBD of LvALF8 counterpart in F. chinensis. However, LvALF8-LBD and FcALF8-LBD exhibited apparently different antibacterial activity against V. parahaemolyticus, the main pathogen causing acute hepatopancreatic necrosis disease (AHPND) of affected shrimp. A structural analysis showed that the positive net charge and amphipathicity characteristics of LvALF8-LBD peptide were speculated as two important components for its enhanced antimicrobial activity compared to those of FcALF8-LBD. These new findings may not only provide some evidence to explain the distinct disease resistance among different shrimp species, but also lay out new research ground for the testing and development of LBD-originated antimicrobial peptides to control of shrimp diseases.     

Molecular Characterization of the Von Willebrand Factor Type D Domain of Vitellogenin from Takifugu flavidus

The von Willebrand factor type D (VWD) domain in vitellogenin has recently been found to bind tetrodotoxin. The way in which this protein domain associates with tetrodotoxin and participates in transporting tetrodotoxin in vivo remains unclear. A cDNA fragment of the vitellogenin gene containing the VWD domain from pufferfish (Takifugu flavidus) (TfVWD) was cloned. Using in silico structural and docking analyses of the predicted protein, we determined that key amino acids (namely, Val¹¹⁵, ASP¹¹⁶, Val¹¹⁷, and Lys¹²²) in TfVWD mediate its binding to tetrodotoxin, which was supported by in vitro surface plasmon resonance analysis. Moreover, incubating recombinant rTfVWD together with tetrodotoxin attenuated its toxicity in vivo, further supporting protein–toxin binding and indicating associated toxicity-neutralizing effects. Finally, the expression profiling of TfVWD across different tissues and developmental stages indicated that its distribution patterns mirrored those of tetrodotoxin, suggesting that TfVWD may be involved in tetrodotoxin transport in pufferfish. For the first time, this study reveals the amino acids that mediate the binding of TfVWD to tetrodotoxin and provides a basis for further exploration of the molecular mechanisms underlying the enrichment and transfer of tetrodotoxin in pufferfish.     

面向养殖水体分布差异的COD光谱法检测研究

面向养殖水体，传统光谱法对化学需氧量（Chemical Oxygen Demand，COD）检测模型构建的基础：源域（现有样本库）与目标域（检测地水体）间光谱数据独立同分布。但是当源域与目标域分布间存在差异时，由源域得到的低误差模型常在目标域上表现下滑。针对该问题，提出面向UV Vis光谱的域对抗训练网络（DAUVwpNet），将分布不同的源域和目标域数据映射至相同分布的特征空间中，使其在该空间的分布距离尽可能接近，从而在特征空间中对源域训练的目标函数也可以迁移至目标域上，以降低模型在目标域的误差。试验表明：面向同一批测试数据，DAUVwpNet的预测误差为0.78，要低于传统模型的预测误差（0.85）；DAUVwpNet预测值与实测值间相关系数为0.95，要高于传统模型的相关系数（0.89）。表明了该网络能够较好对齐两域特征空间数据分布，降低因分布差异带来的COD检测误差。     

Construction and evaluation of introgression lines and fine mapping of ehd8 from Jinghong common wild rice (Oryza rufipogon)

Heading date is one of the most important traits in rice and regulated by multiple genes. Common wild rice is the ancestor of Asian cultivated rice and harbours abundant genetic diversity. To use wild rice resource in rice breeding, a set of 154 introgression lines (ILs) covering 93% of the genome of Jinghong common wild rice was constructed in the background 'Teqing', using 208 simple sequence repeat markers evenly distributed on 12 chromosomes. Among the ILs, the line JIL64 displayed late heading independent of photoperiod. Genetic analysis using the two F₂ populations crossed ''Teqing'/JIL64 and JIL64/'Teqing' revealed that late flowering was controlled by a recessive gene on chromosome 8 (designated early heading date 8, ehd8), and ehd8 was fine mapped to the 50‐kb region flanked by markers RM22221 and 64Indel4. Sequencing and qRT‐PCR demonstrated that LOC_Os08g01410 and LOC_Os08g01420 were deleted in JIL64 and may be associated with the late heading of Jinghong common wild rice. These findings lay a practical foundation for characterizing ehd8, and the ILs help to mine genes from Jinghong common wild rice.     

跟踪采访在参与式技术开发中的应用--以"滇西北农牧生计改良项目"饲料技术开发为例

为了解决滇西北农牧区的冬季饲料紧缺问题,在滇西北贡山县迪麻洛村进行了外来草种和青贮饲料试验.试验采用的主要方法是跟踪采访.跟踪采访的主要内容是监测试验的进展、农户所使用的传统知识以及农户对技术的评价等.在参与式农户试验的条件下,跟踪采访是参与式试验的有效互补手段.     

Preparation of recombinant PTD-HSP27 and verification of its ability to penetrate the cell membrane of human lens epithelial cells and rabbit cornea

AIM: To construct the prokaryotic expression system containing protein transduction domain (PTD) with heat shock protein 27 (HSP27) in order to prepare and purify the recombinant protein, and to verify whether the recombinant protein PTD-HSP27 has the ability to penetrate the human lens epithelial cell (HLEC) membrane and the rabbit cornea. METHODS: The plasmid pKYB-PTD-HSPB1-6His was constructed by the technique of overlap extension PCR. The plasmid was transformed and PTD-HSP27 was purified through nickel affinity chromatography column and identified by Western blotting. PTD-HSP27-6His was labeled with the fluorescein isothiocyanate (FITC). The penetrating ability of PTD-HSP27 into HLECs and rabbit cornea was tested. RESULTS: The recombinant PTD-HSP27 plasmid was successfully cloned and effectively expressed. The correctness of the recombinant protein PTD-HSP27 was demonstrated. Fluorescence microscopic examination showed that PTD-HSP27-FITC was internalized by HLECs. Fluorescent labeled PTD-HSP27 was then observed in the rabbit aqueous humor. CONCLUSION: The recombined gene PTD-HSPB1 was constructed by overlap extension PCR technique and the PTD-HSP27 fusion protein was prepared and purified by nickel affinity chromatography column. Using the technique of PTD-fusion protein, HSP27 was transduced into HLECs and passed through the cornea.     

不同年代大丽轮枝菌菌株的生物学特性和ISSR分析

 为了探究不同年代的大丽轮枝菌(Verticillium dahliae Kleb.)菌株生物学特性和遗传多样性方面的差异,本文以我国黄河流域和长江流域主产棉区6个省的不同年代大丽轮枝菌菌株为研究对象,观察培养性状,测定致病类型(致病力、落叶型),同时采用ISSR指纹图谱分析其遗传多样性。结果显示,不同年代大丽轮枝菌之间菌丝生长速率无显著差异,但菌核型菌株所占比例有减少趋势;2007~2009年和2017年的菌株全部为落叶型菌株,而1983~2000年的菌株中落叶型菌株仅占28.6%,表明随着年代的推移,黄河流域和长江流域的落叶型菌株所占比例呈上升趋势;不同年代菌株之间致病力存在显著差异,1983~2000年、2007~2009年和2017年的菌株中强致病力类型菌株分别占21.4%、25.0%和38.9%,仅有的5株弱致病力类型菌株均为1983~2000年的菌株;与2000年后的菌株相比,1983~2000年的菌株,Nei′s基因多样性指数为0.205 1,Shannon信息指数为0.299 0,表现出更丰富的多样性,利用NTSYS软件和Structure软件对ISSR指纹图谱进行聚类分析,两种方法均将所有供试菌株分为4个类群,且聚类结果与致病力和不同年代之间均具有一定的相关性,与地理来源无明显相关性。本研究结果表明,过去30年间我国黄河流域和长江流域棉田黄萎病菌落叶型和强致病力类型菌株所占比例逐渐升高,且不同年代和不同致病力的菌株在遗传上有差异,为进一步探究大丽轮枝菌的遗传与进化奠定了基础。