欧洲葡萄中CIPK基因的克隆及表达分析

以欧洲葡萄‘佳丽酿’(Carinena)为材料,利用RT-PCR方法获得两个CIPK基因的全长cDNA序列,分别为VvCIPK13和VvCIPK14。VvCIPK13全长为1 501 bp,包括一个1 395 bp的ORF,编码465个氨基酸;VvCIPK14全长为1 616 bp,包括一个1 404 bp的ORF,编码468个氨基酸。生物信息学分析表明两个蛋白均含有两个保守结构域:丝氨酸/苏氨酸激酶结构域和NAF结构域,两个结构域之间有连接域,但VvCIPK13和VvCIPK14的同源性较低,仅有40.33%。利用荧光定量PCR技术研究VvCIPK13和VvCIPK14在葡萄不同组织、植物生长调节剂诱导和逆境胁迫下的表达模式,结果表明:VvCIPK13和VvCIPK14表达具有组织特异性,均在卷须中高表达;VvCIPK13对6-BA诱导有响应,而VvCIPK14对6-BA、ABA、IAA、SA、MeJA和GA_3等诱导均有不同程度的响应;VvCIPK13和VvCIPK14均响应高盐和干旱,但不响应低温,其中VvCIPK14的响应最为强烈。推测VvCIPK13和VvCIPK14在葡萄的非生物胁迫过程中发挥重要的作用。     

Effects of CENP-W down-regulation on human glioma U87 cells

AIM: To study the effect of centromere protein W (CENP-W) down-regulation on human glioma U87 cells.METHODS: Small interfering RNA (siRNA) was used to inhibit the expression of CENP-W in the U87 cells. The interference effect of siRNA was evaluated by RT-qPCR and Western blot. The proliferation of the cells was analyzed by MTT assay, BrdU staining and colony formation experiment. Transwell chamber assay was used to detect the invasion ability of the cells. The cell migration ability was measured by a scratch test. The changes of the cell cycle distribution and apoptosis were analyzed by flow cytometry.RESULTS: The results of MTT assay, colony formation experiment and BrdU staining showed that the cell proliferation and colony formation abilities in experimental group were significantly lower than those in control group and negative control group. The results of Transwell and scratch experiments showed that the migration and invasion abilities in experimental group were weaker than those in blank control group and negative control group. The results of flow cytometry analysis showed that the cell cycle distribution in experimental group was arrested in G₀/G₁ phase. The percentage of apoptotic cells in experimental group was higher than that in control group (P<0.05).CONCLUSION: Down-regulation of CENP-W expression inhibits the proliferation, migration and invasion of human glioma cells and promotes the apoptosis of the cells, suggesting that CENP-W may be a potential target of gene therapy for human glioma.     

Screening of lentiviral vectors carrying effective siRNA targeting S1P2 gene

AIM:To screen the lentiviral vector carrying siRNA with higher efficiency of suppressing the sphingosine-1-phosphate receptor 2(S1P2) gene expression in the primarily cultured corpus cavernosum smooth muscle cells of spontaneously hypertensive rats (SHR).METHODS:SHR and SD rats (n=5 each) were used for primarily culturing corpus cavernosum smooth muscle cells.The cells were randomly divided into 6 groups:SHR siRNA-1,SHR siRNA-2,SHR siRNA-3,SHR GFP,SHR control (SHR non-transfection group),and SD control (SD rat control group).Each group had 5 samples with 1.0×10⁵ cells of each sample.At 72 h after transfection (MOI=60) with lentiviral vectors carrying S1P2 siRNA into the SHR corpus cavernosum smooth muscle cells,the expression of GFP was observed under fluorescence microscope.The protein expression of S1P2,ROCK1,ROCK2 and eNOS in the corpus cavernosum smooth muscle cells,and the mRNA expression of S1P2,ROCK1 and ROCK2 were determined by by Western blot and RT-PCR.RESULTS:The transfection efficiency of the corpus cavernosum smooth muscle cells in SHR siRNA-1,SHR siRNA-2,SHR siRNA-3 and SHR GFP groups were>80%.Compared with SHR control group,the mRNA levels and the protein expression of S1P2,ROCK1 and ROCK2 in SHR GFP group showed no remarkable changes,while those in SHR siRNA-1,SHR siRNA-2,SHR siRNA-3 and SD control groups were significantly lower than those in SHR control group (P<0.05).The protein expression of eNOS in SHR siRNA-1,SHR siRNA-2,SHR siRNA-3 and SHR GFP groups were not significantly changed as compared with SHR control group,but that in SD control group was significantly higher than that in SHR control group.CONCLUSION:Three groups of siRNA lentiviral vectors targeting S1P2 inhibit the expression of S1P2 in the corpus cavernosum smooth muscle cells of SHR,and by silencing the S1P2 expression,the expression of ROCK1 and ROCK2 is inhibited.Among them,siRNA-1 has the highest inhibitory efficiency.     

甜瓜白粉病抗病基因的定位及候选基因分析

以甜瓜高抗白粉病自交系MR-1为母本,易感白粉病自交系Top Mark为父本,构建BC_1P_2和F_2群体,经13个国际通用的甜瓜白粉病生理小种鉴别寄主鉴定,2015年东北农业大学设施园艺工程中心的白粉病发病病菌为P.xanthii生理小种1,甜瓜MR-1对P.xanthii生理小种1的抗性由单显性基因控制。通过对266个F2分离群体的抗病性鉴定和CAPS标记分析,采用复合区间作图法对甜瓜抗白粉病性状进行QTL分析,最终构建了1张包含203个CAPS标记的甜瓜遗传连锁图谱,并将抗病基因PXR定位在第12号染色体上M12-GH和M12-TE 2个标记之间,该基因与两侧翼标记间的距离分别为0.63 c M和0.42 c M,两侧翼标记间在甜瓜参考基因组上对应的物理距离为303 kb,该区域内含有60个预测基因,其中10个基因含有非同义单碱基突变。10个基因荧光定量分析结果显示,在抗病与感病甜瓜表达量存在较大差异的4个基因MELO3C002434、MELO3C002437、MELO3C002441、MELO3C002457为甜瓜白粉病抗病候选基因。     

海拔高度对茅莓果实品质的影响

对梵净山同一山体、同一坡面上800~2 000m不同海拔高度2年生茅莓果实品质进行了分析研究,以期为高海拔地区茅莓栽培选择适宜的生态区提供参考依据。结果表明:可滴定酸含量与海拔高度呈负相关;单果质量、可溶性固形物含量、固酸比、维生素C含量、可溶性蛋白质含量在海拔800~1 800 m区段,随着海拔的升高逐渐增加;而在1 800~2 000m区段,呈下降趋势;在海拔1 600~1 800m区段茅莓的单果质量、可溶性固形物含量、固酸比、维生素C含量、可溶性蛋白质含量与其它海拔区段差异显著(P0.05),表现出很好的品质特性,是适合茅莓优质高产栽培的海拔地带。     

石膏添加量对平菇熟料栽培的影响

以平菇"新831"为试材,采用棉籽壳熟料栽培的方法,研究0%、1%、2%、3%、4%、5%6种石膏添加量对平菇生长发育、污染率和生物学效率的影响,从而确定平菇熟料栽培的最适石膏添加量。结果表明:菌丝长势随石膏添加量增加而减弱;菌丝长速随石膏添加量增加呈先升后降的趋势,石膏添加量为3%时,菌丝长速最快;添加石膏可显著降低菌袋污染率;平菇生物学效率随石膏添加量增加呈先升后降趋势,石膏添加量为1%时,平菇生物学效率最高。综合各项指标得出,棉籽壳熟料栽培平菇石膏的最适添加量为1%。     

我国本地假俭草的特性与应用

在简要介绍假俭草的生态分布及生境类型的基础上,详细介绍了假俭草的耐荫、抗病、抗虫、耐践踏、耐瘠薄、耐旱、耐寒、耐药、抗污染等特性,并与其它暖季型草种进行了比较。回顾了假俭草遗传多样性研究、育种等方面的进展及应用情况。提出假俭草新品种可实现草坪的"少修剪、不打药、不施肥、不灌溉"的低养护管理目标,其推广应用可最大限度地降低草坪的养护成本,为我国城市绿地及球场建设做出贡献。建议在广泛收集全国野生假俭草种质资源的基础上,加强新品种选育工作,培育优良品系,加大引种繁育力度,扩大其适应地域,以提高假俭草利用率,促进我国草业的发展。     

套袋苹果影像的分割方法研究

为了准确识别并分割套袋苹果,对影像中的套袋苹果及其背景在RGB颜色空间模式下,分析各目标物的R(红)、G(绿)及B(蓝)值分布特征,根据各目标物所呈现的R、G及B值差异选取适合于套袋苹果的分割条件,对套袋苹果进行分割,将分割结果与多阈值分割法以及K均值聚类法分割结果进行比较。结果表明:分割法优于以上2种方法,同时,根据该试验提出的分割方法计算苹果识别率,识别率可达93.4%,表明该试验算法对套袋苹果的分割准确率高。     

新疆加工型辣椒花药培养技术研究

以6个新疆加工型辣椒品种207、208、64、66、67、HT113为试材,以MSB培养基为基本培养基,附加不同植物激素浓度的NAA、2,4-D、TDZ、KT进行花药培养,研究不同植物激素处理对加工型辣椒花药愈伤组织和胚状体诱导的影响,以建立适应于新疆加工型辣椒的花药培养技术。结果表明:不同基因型辣椒的愈伤组织诱导率和出胚率均不同,愈伤组织诱导率为4.98%~29.65%,胚状体诱导率为0~6.88%。0.5 mg·L~(-1) 2,4-D+0.1mg·L~(-1) TDZ和0.5mg·L~(-1) NAA+0.1mg·L~(-1) KT分别是愈伤组织诱导和胚状体诱导最适宜的激素组合;在新疆特有的气候条件和种植模式下,7月中旬前的花药接种培养最为适宜;胚状体继续发育成熟后,通过练苗移栽获得了56株单倍体植株。     

大棚苦瓜白粉病田间药效试验

开展了四氟醚唑、醚菌酯、丙环·醚菌酯、甲基硫菌灵4种药剂防治大棚苦瓜白粉病的田间药效试验。试验结果表明,4种药剂均可有效控制大棚苦瓜白粉病发展,且对苦瓜安全无药害,以32%丙环·嘧菌酯悬浮剂1 500倍液防效最好,其次是4%四氟醚唑水乳剂1 000倍液、50%醚菌酯水分散粒剂2 000倍液、70%甲基硫菌灵可湿性粉剂600倍液,在生产上防治时可交替使用。