白屈菜CmFAD2基因的克隆及植物转化载体构建

通过对白屈菜低温应答过程的转录组分析发现膜脂不饱和化相关基因的表达在一定过程中发生变化,脂肪酸去饱和酶基因FAD2在随温度的变化趋势为正"V"型,且表达量变化显著。利用NCBI等在线软件对序列进行相关生物学信息分析,并对白屈菜FAD家族成员FAD2基因的完整开放阅读框(ORF)进行克隆,并命名为CmFAD2。选用克隆载体pMD-19-T,转化大肠杆菌DH5α,测序验证序列正确性及完整性。将目的基因与植物表达载体pRI-201-AN连接构建重组DNA pRI-201-AN-Cm FAD2,电击法转化农杆菌LBA4404,利用菌液PCR法验证成功。该基因可作为药用植物抗寒品种创制的候选基因。     

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。     

正交设计优化青蒿SRAP-PCR反应体系及引物筛选

为了建立青蒿的SRAP最佳扩增体系,并筛选出SRAP多态性引物,本研究以青蒿叶片DNA为模板,采用正交试验设计,以Mg^2+、dNTP Mix、Taq DNA聚合酶、引物和DNA模板5种因素5个水平,对青蒿SRAP反应体系进行研究。结果表明,青蒿SRAP-PCR最佳反应体系为:引物0.6μmol/L、Mg^2+2.0 mmol/L、模板DNA 5.1 ng、Taq DNA聚合酶2.0 U、dNTPs 0.25 mmol/L,总体积为25μL。各因素对扩增反应均有不同影响,其中引物浓度的影响最大,dNTPs的影响最小。运用该体系对不同种质资源的青蒿进行验证,证明该体系稳定可靠,并在30个引物组合中筛选出了25对扩增条带清晰,多态性丰富的引物组合。这一结论为今后利用SRAP标记技术进行青蒿分子遗传学研究提供了科学依据。     

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。     

槟榔间作条件下不同修剪方式对可可生长和产量的影响

以槟榔间作可可条件下的可可为研究对象,比较不同修剪方式对其生长、光合作用及产量的影响。结果表明:中度修剪提高可可株高、干周和主枝粗度,重度修剪和仅控冠幅降低可可干周和主枝粗度。中等修剪可可叶片Pn显著高于轻度修剪和仅控冠幅,可能由于修剪提高了新梢叶片叶肉细胞光合作用能力。中度修剪可可徒长枝重量分别比轻度修剪、重度修剪和仅控冠幅显著降低了69.27%、54.69%和57.03%,大大降低了养分消耗,提高产量的同时节省了修剪工作量。综上所述,在槟榔间作可可条件下,采用中度修剪有利于提高光合作用效率,形成健康平衡树形,降低徒长枝养分消耗,提高产量,为槟榔间作可可节本增效技术研究与优化提供依据。     

Identification,characterization and full-length sequence analysis of a novel endornavirus in common sunflower (Helianthus annuus L.)

To identify the possible quarantine viruses in seven common sunflower varieties imported from the United States of America and the Netherlands, we tested total RNAs extracted from the leaf tissues using next-generation sequencing of small RNAs. After analysis of small RNA sequencing data, no any quarantine virus was found, but a double-stranded RNA(dsRNA) molecule showing typical genomic features of endornavirus was detected in two varieties, X3939 and SH1108. Full-length sequence and phylogenetic analysis showed that it is a novel endornavirus, temporarily named as Helianthus annuus alphaendornavirus(HaEV). Its full genome corresponds to a 14 662-bp dsRNA segment, including a 21-nt 5′ untranslated region(UTR), 3' UTR ending with the unique sequence CCCCCCCC and lacking a poly(A) tail. An open reading frame(ORF) that encodes a deduced 4 867 amino acids(aa) polyprotein with three domains: RdRP, Hel and UGT(UDP-glycosyltransferase). HaEV mainly distributed in the cytoplasm but less in the nucleus of leaf cells by fluorescence in situ hybridization(FISH) experiment. This virus has a high seed infection rate in the five varieties, X3907, X3939, A231, SH1108 and SR1320. To our knowledge, this is the first report about the virus of the family Endornaviridae in the common sunflower.     

Establishing an eyeball-weight relationship for Litopenaeus vannamei using machine vision technology

This study aimed to establish a shrimp eyeball-weight relationship model for Litopenaeus vannamei using machine vision technology. A total of 295 shrimp were sampled from a recirculating aquaculture system (RAS). The long-axis length (d), body length (L), and body weight (W) of each individual was measured. The long axis length of the shrimp eyeball was identified and measured using machine vision technology. Continuous fitting and piecewise fitting models were used to construct the eyeball-weight relationship model for L. vannamei. The continuous fitting relationship model was described as: W = 38.865d^2.7914, while the piecewise model was described as: d < 2 mm, W = 0.0326d^3.7363, R² = 0.9288; 2 mm ≤ d < 3.9 mm, W = 0.0401d^3.104, R² = 0.9629; 3.9 mm ≤ d < 5.8 mm, W = 0.0421d^3.0311, R² = 0.9216; 5.8 mm < d, W = 0.103d^2.6226, R² = 0.9457. The root mean square error (RMSE) of the piecewise fitting model (0.0244, 0.1575, 0.5034, 0.7072) was smaller than the continuous fitting model (0.8229). The correlation coefficient (R²) of the piecewise model (0.9288, 0.9629, 0.9216, and 0.9457) was similar to that of the continuous fitting model (R² = 0.9621). The results indicated that the piecewise fitting model is suitable for calculating the biomass of L. vannamei in RAS and provides a novel way of estimating the biomass of L. vannamei cultured in RAS. The piecewise fitting model can also provide the foundation of evaluating the production of shrimp using underwater image recognition in intelligent aquaculture systems.     

同源四倍体及其原二倍体黑皮冬瓜光合光响应模型筛选

为了筛选出最适合黑皮冬瓜Benincasa hispida (Thunb.) Cogn.的光合光响应模型,为其育种提供理论依据,以同源四倍体及其原二倍体黑皮冬瓜为试验材料,对8种经典的光合光响应模型适用性进行了比较分析。结果表明：二次多项式能够表现出光抑制情况,但在拟合过程中出现暗呼吸速率为正值、光补偿点为负值及无法解释当光强达到饱和后光合速率快速下降的问题;直角双曲线、非直角双曲线及指数函数Ⅰ、指数函数Ⅱ无法直接求取光饱和点、光补偿点,结合常用的光饱和点的计算方法得到的光饱和点与实测值均存在较大的偏差,且指数函数Ⅱ在计算光饱和点时表现出明显的人为性,也无法拟合光抑制情况,但4种模型拟合得到的光补偿点均与实测值相差不大;指数修正模型因系数β为负值,无法求取四倍体黑皮冬瓜材料的光饱和点和最大净光合速率,且拟合得到的四倍体黑皮冬瓜的光补偿点明显低于实测值;直角双曲线修正模型计算得到的暗呼吸速率及二倍体黑皮冬瓜的光饱和点明显低于实测值,但获得的四倍体及其二倍体的最大净光合速率与实测值最接近,说明其在拟合最大净光合速率上有优势;整体上分段函数计算得到的黑皮冬瓜的各光合参数与实测值最为接近,与实测值的平均相对误差最小,也能很好的拟合发生光抑制部分的光响应曲线。分段函数拟合同源四倍体及其原二倍体黑皮冬瓜光合光响应曲线效果较其他模型效果好,分段函数模型为黑皮冬瓜最适合的光合光响应模型。     

围场县马铃薯中典型药物污染特征及健康风险评价

为掌握典型药物在农作物中的污染特征及健康风险,保障农产品质量安全,利用超声萃取-固相萃取-高效液相色谱/串联质谱(LC-MS/MS)技术,对围场县全县马铃薯中5大类(磺胺类、四环素类、氟喹诺酮类、大环内酯类及其他类)25种药物污染水平及富集效果进行了调查,并对健康风险进行了评价。结果表明,有20种目标物被检出,不同目标物检出率差异显著,检出含量范围为0.13～3.67μg·kg-1,单种污染物最高检出含量为17.0μg·kg-1。马铃薯对目标污染物的富集系数范围在0.01～39.6之间,最大富集系数为61.8。畜禽粪便是马铃薯中药物残留的主要来源,尽管整体检出水平和健康风险相对较低,但长期摄入与协同作用引发的潜在健康风险不容忽视。因此,仍需严控污染源头,推进畜禽粪便安全资源化利用,确保农田环境质量及农产品安全品质。