巴西固氮螺菌Yu62 glnB基因和glnZ基因的克隆和序列分析

ｇｌｎＢ基因编码的ＰⅠⅠ蛋白在巴西固氮螺菌的固氮过程中起着非常重要的调控作用,而ｇｌｎＺ是与ｇｌｎＢ高度同源的基因,其产物ＰＺ可能也在固氮调控中起作用。本研究用ＰＣＲ法克隆了巴西固氮螺菌Ｙｕ６２ ｇｌｎＢ基因和ｇｌｎＺ基因。ＤＮＡ序列分析表明ｇｌｎＢ和ｇｌｎＺ这２个基因的编码区的长度都为３３６ｂｐ,编码１１２个氨基酸。将巴西固氮螺菌Ｙｕ６２菌株与标准菌株Ｓｐ７的ｇｌｎＢ基因和ｇｌｎＺ基因分别进行比     

高产、多抗、紧凑、中早熟玉米杂交种强盛62的选育

强盛62是由山西省农科院食用菌研究所玉米育种部于2003年以自选系3412为母本,9248为父本杂交育成的紧凑、中早熟玉米杂交种。经过2007～2008年山西省早熟玉米区试试验和生产试验,表现出高产、抗病抗倒、早熟等特点,在生产上具有较高的推广应用价值。     

稻瘟病菌假定的糖基水解酶62家族初步研究

 【目的】了解糖基水解酶62家族细胞壁降解酶系在稻瘟病菌致病中的作用。【方法】通过生物信息学方法对稻瘟病菌基因组中假定的糖基水解酶62家族成员进行基因结构、蛋白分泌特性及系统发育分析,并对其中一个成员MGG_01403.6进行过量表达、基因敲除分析。【结果】该家族共有8个成员,均具有细胞壁降解酶（糖基水解酶62家族）保守结构域,且均为胞外分泌蛋白;系统发育分析可以将这些成员分别聚类在两个进化分支中;成员间在不同侵染阶段表达模式有差异;MGG_01403.6过量表达和基因敲除均不影响稻瘟病菌的致病性。【结论】糖基水解酶62家族可能存在功能冗余作用,进一步的双突变或多突变可能是明确这类多基因家族基因功能的重要方法。     

播期对阴山北麓干旱半干旱区华油杂62生长及养分吸收的影响

以饲料油菜华油杂62为研究对象,在阴山北麓干旱半干旱农牧交错区,通过分期播种,研究了播期对饲料油菜生长发育、产量及养分吸收累积的影响。结果表明:播期推迟导致饲料油菜生育期缩短,尤其缩短了从播种到出苗的时间;有效分枝数、茎粗随播期推迟而增大;出苗数、鲜草产量、养分累积量随播期推迟而下降;但植株地上部N、P、K含量随播期推迟变化趋势各不相同,氮含量先减小后增大,磷含量先增大后降低,钾含量逐渐降低;随播期推迟氮、钾累积量逐渐降低,磷积累量先增大后降低。综合考虑,阴山北麓干旱半干旱农牧交错区饲料油菜最适宜播期应为5月下旬至6月上旬。     

巴西橡胶树HbAGL62基因的克隆及表达分析

从巴西橡胶树早花材料与普通种质cDNA差减文库中筛选到一个具有MADS保守序列的EST基因片段,根据该片段序列信息设计特异引物,利用RACE进行特异片段的5′扩增,获得长度为935 bp的HbAGL62基因克隆。生物信息学分析表明,该基因包含795 bp的开放阅读框,编码265个氨基酸,含有MADS保守结构域,与拟南芥、葡萄和苜蓿AGL62基因进化距离最近。实时荧光定量PCR表明,HbAGL62在橡胶树根和皮中不表达,在花和胚中表达,存在组织特异性表达;在叶芽分化阶段不表达,花芽分化时高效表达,表明HbAGL62与橡胶树开花调控有重要关系。     

Cytochrome P-450 mediated O-dealkylation of 7-alkoxycoumarins in liver microsomes from rainbow trout (Salmo gairdneri)

Evidence has recently been presented for variation in the inducibility of various 7-alkoxycoumarin-O-dealkylase activities in liver microsomes from a number of mammalian species by -naphthoflavone (NF). In the present study we have investigated the inducibility of hepatic microsomal 7-methoxycoumarin-O-demethylase, 7-ethoxycoumarin-O-deethylase, 7-propoxycoumarin-O-depropylase and 7-butoxycoumarin-O-debutylase activities in rainbow trout by NF. O-demethylase activity was increased approximately 17-fold, O-deethylase and O-depropylase activities approximately 9-fold and O-debutylase activity approximately 25-fold. The kinetics of the various hepatic microsomal 7-alkoxycoumarin-O-dealkylase activities were investigated in control and NF-treated rainbow trout. The O-demethylase-, O-depropylase- and O-debutylase activities exhibited monophasic Michaelis-Menten kinetics in liver microsomes from both control and NF-treated rainbow trout, whereas the O-deethylase activity exhibited biphasic Michaelis-Menten kinetics in control liver microsomes and monophasic Michaelis-Menten kinetics in liver microsomes from NF-treated rainbow trout.     

四季秋海棠BsMYB62的抗旱性功能研究

【目的】以前期从四季秋海棠叶片中克隆得到的R2R3型MYB转录因子BsMYB62为研究对象,分析其在干旱胁迫下的功能。【方法】对四季秋海棠MYB62蛋白进行亚细胞定位,并通过实时荧光定量PCR方法明确BsMYB62基因在四季秋海棠根、茎、叶、雄花和雌花5个组织部位中的表达情况;构建BsMYB62基因的过表达载体转化到拟南芥,观察过表达转基因株系与野生型植株在萌发期和幼苗期遭受干旱胁迫时的生长表型、种子萌发和根长情况,测定各株系在幼苗期干旱胁迫处理时的叶绿素荧光参数PSⅡ最大光化学效率（F_v/F_m）,叶绿素、丙二醛（MDA）、脯氨酸（Pro）含量,抗氧化酶（SOD、POD、CAT）活性以及转基因株系中BsMYB62基因的相对表达量变化。【结果】BsMYB62定位于四季秋海棠的细胞核,其在根、茎、叶和雌雄花中均有表达,其中以根和茎中的表达水平较高。在MS培养基中,野生型拟南芥和3个BsMYB62过表达转基因株系（OE1、OE2和OE3）的发芽率和长势均无显著差异,而在200 mmol/L甘露醇（模拟干旱胁迫）培养基中,OE1、OE2和OE3在胁迫处理后前2 d的发芽率及主根长均显著高于野生型拟南芥,幼苗长势也明显优于野生型。置于培养土中干旱胁迫1 d时,OE1、OE2和OE3的BsMYB62即被显著诱导;干旱胁迫13 d时,OE1、OE2和OE3的F_v/F_m、叶绿素含量、Pro含量以及SOD、POD和CAT活性均显著高于野生型拟南芥,而MDA含量则显著低于野生型拟南芥。【结论】BsMYB62基因通过提高转基因拟南芥的光合潜能和抗氧化能力,显著增强其对干旱胁迫的抗性。     

Injury effect of adenosine triphosphate on N9 microglia

AIM:To investigate the injury effect of adenosine triphosphate(ATP) on N9 microglia. METHODS:N9 microglia in logarithmic growth phase was randomly divided into 3 groups. In control group, the cells were cultured without ATP treatment. In ATP group, the cells were treatment with ATP after cultured for 24 h. In KN-62 intervention group, after pretreatment with KN-62 for 30 min, ATP was added in the cells. The cell viability was assessed by XTT assay. Cellular morphological changes were observed under phase-contrast microscope. The cell cycle and apoptosis were detected by flow cytometry. The expression of P2X₇ receptor was examined by immunofluorescence staining. The protein levels of P2X₇ receptor were measured by Western blotting. The concentration of IL-1β in the culture supernatant was detected by ELISA. RESULTS:ATP at dose of 500 μmol/L and 1 mmol/L only caused small damage to the cell viability of N9 microglia. The cell viability was 88.5%±5.5% and 88.2%±8.4% after treated with ATP for 24 h,respectively. The cell viability dropped rapidly and cell shrinkage occurred when the concentration of ATP increased to 2 mmol/L or higher. With the extension of experiment time, the cell viability and cell density decreased further and cell shrinkage was getting worse. KN-62 intervention improved the viability of N9 microglia injured by ATP. The morphology and density of N9 microglia in KN-62 intervention group were much better than those in ATP group. ATP arrested N9 microglia at S phase and increased cell apoptosis significantly(P<0.01 vs control group). KN-62 intervention obviously relieved the cell cycle arrest and decreased the cell apoptosis caused by ATP(P<0.01). ATP and KN-62 intervention had no effect on the distribution of P2X₇ receptor. The protein levels of P2X₇ receptor had no significant difference among the 3 groups(P>0.05). ATP and KN-62 intervention had no effect on the release of IL-1β. CONCLUSION:High dose of ATP damages N9 microglia and its mechanism may be related to cell cycle arrest and apoptosis mediated by P2X₇ receptor but not to inflammatory response caused by microglia.     

冬小麦抗条锈新品系95-62-1选育报告

兰州商学院小麦研究所与兰州农业学校、天水农业学校协作 ,以 Ibis为父本、兰天 10为母本育成的冬小麦新品系 95 - 6 2 - 1,在 2 0 0 1- 2 0 0 3年的天水市高山组旱地小麦区域试验中折合平均产量 5 35 5 .0 kg/ hm2 ,较对照品种中梁 2 2增产 7.3% ;成株期对条中 2 9、条中 31、条中 32、条中 32 (中梁 17- S)、洛 13 、水 3、水 4、水 14及混合菌全部免疫 ;越冬率为 94 .2 %～ 10 0 % ;经甘肃省农科院测试中心检测 ,其籽粒含粗蛋白 15 .83%、赖氨酸 0 .5 1%。主要适宜在陇南高山区及二阴半山区种植     

Changes in some blood micronutrients, leukocytes and neutrophil expression of adhesion molecules in periparturient dairy cows.

Dairy cows are highly susceptible to infectious diseases, like mastitis, during the period around calving. Although factors contributing to increased susceptibility to infection have not been fully elucidated, impaired neutrophil recruitment to the site of infection and changes in the concentrations of some micronutrients related with the function of the immune defence has been implicated. Most of the current information is based on studies outside the Nordic countries where the conditions for dairy cows are different. Therefore, the aim of the study was to evaluate changes in blood concentrations of the vitamins A and E, the minerals calcium (Ca), phosphorous (P), and magnesium (Mg), the electrolytes potassium (K) and sodium (Na) and the trace elements selenium (Se), copper (Cu) and zinc (Zn), as well as changes in total and differential white blood cell counts (WBC) and expression of the adhesion molecules CD62L and CD18 on blood neutrophils in Swedish dairy cows during the period around calving. Blood samples were taken from 10 cows one month before expected calving, at calving and one month after calving. The results were mainly in line with reports from other countries. The concentrations of vitamins A and E, and of Zn, Ca and P decreased significantly at calving, while Se, Cu, and Na increased. Leukocytosis was detected at calving, mainly explained by neutrophilia, but also by monocytosis. The numbers of lymphocytes tended to decrease at the same time. The mean fluorescent intensity (MFI) of CD62L and CD18 molecules on blood neutrophils remained constant over time. The proportion of CD62L+ neutrophils decreased significantly at calving. The animals were fed according to, or above, their requirements. Therefore, changes in blood levels of vitamins, minerals and trace elements were mainly in response to colostrum formation, changes in dry matter intake, and ruminal metabolism around calving. Decreased levels of vitamins A and E, and of Zn at calving might have negative implications for the functions of the immune defence. The lower proportion of CD62L+ neutrophils at calving may result in less migration of blood neutrophils into the tissues, and might contribute to the increased susceptibility to infections at this time.