玉米Gln1-3 gDNA序列分离、基因结构、保守功能域与等位变异分析

谷氨酰胺合成酶家族是高等植物体内氨态氮同化酶, 是植物体内氮利用与循环的核心构件。玉米Gln1-3基因编码叶肉细胞中的细胞质同工酶, 与全株氮向籽粒蛋白质同化与氮利用效率及产量紧密相关。本研究以玉米自交系辽白371为材料, 采用PCR与接头步移(walking)方法分离得到Gln1-3基因区域基因组DNA(gDNA)全长4 571 bp, 起始密码子至终止密码子序列长3 062 bp。将该基因在GenBank注册, 登录号为EU338535, 并进行了详细注释。该基因由10个外显子、9个内含子组成, 剪接方式主要为5′供位保守的GU与3′受位AG模式。编码的GS1蛋白由356个氨基酸组成, 分子量39.2 kD, 等电点 (pI) 为5.202。保守功能域分析表明, 氨基末端外显子2到外显子6为氨离子结合结构域, 羧基末端外显子8与外显子9构成ATP酶结构域, 在胞内行使催化功能。对50个玉米自交系Gln1-3基因重要分析区域进行了等位变异分析, 鉴定出364个等位变异, 其中313个为SNP变异, 占86%。     

小麦及大麦幼苗对供锌及添加CaCO3/HCO3-的反应

石灰性土壤含有大量CaCO3所产生的高含量HCO3-及CaCO3对Zn的吸附作用被认为是造成作物缺锌的重要原因.该研究在营养液中添加CaCO3或HCO3-,探讨了两者在不同供Zn条件下对2种小麦(小偃22、郑麦9023)与1种大麦(西引2号)幼苗生长及Zn吸收的影响.结果表明,不供Zn条件下,添加HCO3-时3种供试作物均出现明显失绿症状,供Zn时症状减轻;同时HCO3-明显抑制小麦和大麦生长,供Zn抑制作用减轻.供Zn显著增加幼苗的Zn吸收,且地上部增加幅度高于根系;3种供试作物均对缺锌较为敏感;HCO3-通过抑制幼苗生长及Zn从根系向地上部转运而影响Zn吸收,大麦的锌转运率高于小麦.HCO3-还显著提高了营养液pH,但介质中有作物时pH值增加幅度较小;与HCO3-相比,高量CaCO3对作物生长及Zn吸收影响较小,但能明显提高营养液pH值.     

玫瑰黄链霉菌Men-myco-93-63负调控基因nsdAmgh的克隆及序列分析

nsdA (Negative regulator of Streptomyces differentiation)基因是天蓝色链霉菌中发现的与抗生素合成相关的负调控基因.根据天蓝色链霉菌 A3(2)的序列设计引物扩增玫瑰黄链霉菌 Men-myco-93-63 菌株中的该基因,经测序和序列分析表明,由保守序列引物nsdA-2R和nsdA-2L克隆到的Men-myco-93-63 菌株中的该基因 800 bp片段与已知nsdA基因核苷酸保守区域序列同源性达到99%,由全长序列扩增引物nsdA WZ-R和nsdA WZ-L克隆到的该基因包含一个完整的开放阅读框,共编码 369 个氨基酸,与变铅青链霉菌和天蓝色链霉菌A3(2)中的nsdA 基因的核苷酸序列同源性达到100%,氨基酸序列同源性达到99%.该全长基因命名为nsdA_(mgh),其在玫瑰黄链霉菌 Men-myco-93-63 中的发现为阻断该负调控基因以期获得抗生素高产工程菌株提供了重要依据.     

玉米种质C8605-2在品种改良中的应用

C8605-2是国内优良的玉米骨干自交系之一,它在新品种选育和种质改良创新中都得到了广泛的应用。加强其研究和利用,对丰富和发展种质资源、培育优良品种必将会起到极大的促进作用。     

Regeneration of adventitious shoots from mature stored cotyledons of Japanese plum (Prunus salicina Lind1)

Adventitious shoot regeneration from mature stored cotyledons of Japanese plum (Prunus salicina Lind1) was achieved in vitro. The influences of the presence and absence of the light, different concentrations of thidiazuron (TDZ) and benzyladenine (BA) in the culture media, TDZ pretreatments and different basal salts on shoot regeneration were evaluated. TDZ was more effective in inducing shoot regeneration from mature stored cotyledons than BA. Dark incubation significantly increased the regeneration frequencies. Quoirin/Lepoivre (QL) basal salts stimulated shoot regeneration more than woody plant (WPM) or B5 salts did. The frequency of adventitious shoot formation varied among the varieties and the regeneration ability appeared to be genotype depended. The frequency of regeneration under the optimum tested conditions for ‘Bruce’, ‘Shiro’, ‘Redheart’, ‘Gladstone’ and ‘Early Golden’ cotyledons were 66.7%, 46.7%, 43.3%, 26.7% and 6.7%, respectively.     

3个豇豆株系的性状观察比较试验

对豇豆99-2,99-3,99-4三个优良株系性状进行比较试验.结果表明,豇豆99-2具有生长势好、早熟、高产、优质、商品性好等特点,丰产性尤为突出;前期产量12 304.5 kg/hm2,占总产量33 949.5 kg/hm2的36.68%,鲜荚比99-3、99-4早2～3 d上市.可作为早熟品种推广栽培.     

Nitrate leaching in a silage maize field under different irrigation and nitrogen fertilizer rates

Quantification of the interactive effects of nitrogen (N) and water on nitrate (NO₃) loss provides an important insight for more effective N and water management. The goal of this study was to evaluate the effect of different irrigation and nitrogen fertilizer levels on nitrate-nitrogen (NO₃-N) leaching in a silage maize field. The experiment included four irrigation levels (0.7, 0.85, 1.0, and 1.13 of soil moisture depletion, SMD) and three N fertilization levels (0, 142, and 189 kg N ha⁻¹), with three replications. Ceramic suction cups were used to extract soil solution at 30 and 60 cm soil depths for all 36 experimental plots. Soil NO₃-N content of 0-30 and 30-60-cm layers were evaluated at planting and harvest maturity. Total N uptake (NU) by the crop was also determined. Maximum NO₃-N leaching out of the 60-cm soil layer was 8.43 kg N ha⁻¹, for the 142 kg N ha⁻¹ and over irrigation (1.13 SMD) treatment. The minimum and maximum seasonal average NO₃ concentration at the 60 cm depth was 46 and 138 mg l⁻¹, respectively. Based on our findings, it is possible to control NO₃ leaching out of the root zone during the growing season with a proper combination of irrigation and fertilizer management.     

稀土壳聚糖螯合盐对水产颗粒饲料性能的影响

研究了在饲料中添加稀土壳聚糖螯合盐（ＲＥＣＣ）对水产颗粒饲料性能的影响。根据鲫（Ｃａｒａｓｓｉｕｓａｕｒａｔｕｓ）鱼苗的营养需求配制４种类型的ＲＥＣＣ试验日粮,即１号饲料（０．００％）、２号饲料（０．０８％）、３号饲料（０．１６％）和４号饲料（０２４％）。测定４种饲料的粉化率和溶失率。结果显示,添加稀土壳聚糖螯合盐后可以降低饲料的 粉化率,饲料中添加０．１６％的稀土壳聚糖螯合盐效果尤其明显（Ｐ＜０．０１）;在饲料中添加ＲＥＣＣ可以显著降低饲料的溶失率（Ｐ＜０．０５）,并可以提高和改善水产颗粒饲料的性能。     

MicroRNA-142-3p modulates atherosclerosis-associated endothelial cell apoptosis via targeting Rictor

AIM To investigate the role of microRNA-142-3p （miR-142-3p） in endothelial cell apoptosis during atherosclerosis （AS） and the underlying mechanism. METHODS Human aortic endothelial cells （HAECs） were treated with oxidized low-density lipoprotein （ox-LDL）. The expression level of miR-142-3p was detected by RT-qPCR. Apoptosis was determined via flow cytometry （FCM） and caspase-3 activity assay. Prediction of the binding site between miR-142-3p and 3’-UTR of Rictor mRNA was performed by bioinformatics analysis and confirmed by dual-luciferase reporter assay. RESULTS The expression of miR-142-3p was substantially up-regulated during the ox-LDL-elicited apoptosis in HAECs （P<0.05,P<0.01）. Forced expression of miR-142-3p exacerbated apoptosis in HAECs whereas inhibition of miR-142-3p partly alleviated apoptotic cell death mediated by ox-LDL. Further analysis identified Rictor as a direct target gene of miR-142-3p, and Rictor knock-down abolished the anti-apoptotic effect of miR-142-3p inhibitor. Moreover, the Akt/endothelial nitric oxide synthase （eNOS） signaling pathway was found to mediate the beneficial effect of miR-142-3p inhibitor on endothelial cells apoptosis. CONCLUSION Down-regulation of miR-142-3p inhibits endothelial cell apoptosis and atherosclerotic development by up-regulating the expression of Rictor and activating the Akt/eNOS signaling pathway.     

Effect of lncRNA FEZF1-AS1 on viability and apoptosis of LPS-induced vascular endothelial cells by regulating miR-363-3p expression

AIM To investigate the mechanism of long noncoding RNA （lncRNA） FEZF1-AS1 regulating microRNA-363-3p （miR-363-3p） on the viability and apoptosis of lipopolysaocharide （LPS）-induced vascular endothelial cells. METHODS Human umbilical vein endothelial cells （HUVECs） were cultured in vitro. pcDNA-NC, pcDNA-FEZF1-AS1, anti-miR-NC, anti-miR-363-3p, miR-NC and miR-363-3p mimics were transfected into the HUVECs and LPS stimulation was applied for 24 h. RT-qPCR was used to detect the expression of FEZF1-AS1 and miR-363-3p. The cell viability was measured by MTT assay. The apoptotic rate was analyzed by flow cytometry. The dual-luciferase reporter experiment was used to verify the targeted regulation of FEZF1-AS1 and miR-363-3p. Western blot was used to determined the expression of cyclin D1, Ki67 and cleaved caspase-3. RESULTS Compared with control group, the expression level of FEZF1-AS1 in LPS group was significantly reduced （P<0.05）, and the expression level of miR-363-3p was significantly increased （P<0.05）. Compared with pcDNA-NC+LPS group, the cell viability in pcDNA-FEZF1-AS1+LPS group was significantly increased （P<0.05）, the apoptotic rate was significantly reduced （P<0.05）, the protein levels of cyclin D1 and Ki67 were significantly increased （P<0.05）, and the protein level of cleaved caspase-3 was significantly reduced （P<0.05）. Compared with anti-miR-NC+LPS group, the cell viability in anti-miR-363-3p+LPS group was significantly increased （P<0.05）, the apoptotic rate was significantly reduced （P<0.05）, the protein levels of cyclin D1 and Ki67 were significantly increased （P<0.05）, and the protein level of cleaved caspase-3 was significantly reduced （P<0.05）. Dual-luciferase reporter experiment confirmed that FEZF1-AS1 targeted miR-363-3p. Compared with miR-NC+pcDNA-FEZF1-AS1+LPS group, the cell viability in miR-363-3p+pcDNA-FEZF1-AS1+LPS group was significantly reduced （P<0.05）, the apoptotic rate was significantly increased （P<0.05）, the protein levels of cyclin D1 and Ki67 were significantly reduced （P<0.05）, and the protein level of cleaved caspase-3 was significantly increased （P<0.05）. CONCLUSION Over-expression of FEZF1-AS1 promotes the viability and inhibits apoptosis of LPS induced vascular endothelial cells by inhibiting the expression of miR-363-3p.