板栗脂质转运蛋白基因的克隆及表达

利用RACE技术从板栗短雄花序突变体中扩增得到660bp的板栗脂质转运蛋白(lipid transfer protein,LTP)cDNA片段。该cDNA编码118个氨基酸,具有8个位置保守的半胱氨酸(C)残基及26个氨基酸的信号肽,它与棉花、草莓脂质转运蛋白氨基酸序列相似性为63%,基因序列已提交数据库(GenBank),其登录号分别为FJ490676(基因)和ACL01093(蛋白)。荧光定量分析表明板栗短雄花序突变体较正常雄花序的脂质转运蛋白表达量更高。将板栗脂质转运蛋白基因插入到硫氧还蛋白融合表达载体pET32a(+)中,构建了板栗的原核表达载体pET-LTP,在大肠杆菌菌株Rosetta-gamiTM2(DE3)中,IPTG诱导5h大量表达约为30ku的融合蛋白,通过Ni2+-chelating sepharose fast flow柱纯化的融合蛋白具有抑制板栗镰刀菌属病原真菌孢子萌发的功能。     

Differential expression of defence-related proteins in Vigna unguiculata (L. Walp.) seedlings after infection with Fusarium oxysporum

This work aimed to study the induction of defence proteins in cowpea seedlings during the first days after infection with the fungi Fusarium oxysporum f. sp. cubenses and F. oxysporum f. sp. phaseoli. Cowpea seeds, after disinfection, were transferred to Petri dishes containing 0.5% agar and, after germination, were infected with a drop of a suspension containing 0.5 × 10⁴ spores ml⁻¹. Seedlings were collected at 24, 48, 72 and 96 h after infection and were dissected into leaves, hypocotyls, roots, cotyledons and teguments, which were measured and weighed for morphometric analysis. The agar medium was also analyzed. Enzymatic assays of proteic extracts yielded antimicrobial peptides detected by Western blotting. The cowpea seedlings showed a complex pattern of induction and repression of defence proteins in response to infection by both pathogens. Furthermore, morphometric analysis showed differences between infected and control seedlings. Infected samples did not at any time exhibit chitinase activity, but did exhibit different β-1,3-glucanase and peroxidase activities. Western blotting for lipid transfer protein (LTP) demonstrated its presence in all parts of the infected seedlings. Exuded proteins, also obtained from cowpea seeds in the germination medium, were separated by SDS-PAGE and tricine gel electrophoresis. The analysis showed that some proteins were exuded from moistened cowpea seeds, particularly after F. oxysporum infection.     

两种抗病基因双T-DNA区双元载体的构建

选择标记基因的删除是植物转基因工程中重要的步骤之一,双T-DNA区双元载体转化法以其共整合频率高、删除标记基因效率高的优点在植物遗传转化中应用非常广泛。本文通过构建天麻抗真菌蛋白基因、脂质转移蛋白基因双T-DNA区双元载体,为培育不含选择标记基因的抗病新材料奠定基础。通过PCR的方法克隆GAFP基因,产物纯化测序确定序列准确后与pSB130-35S载体进行BamHⅠ、SacⅠ双酶切;pCAMBIA2300-LTP、pSB130-35S经EcoRⅠ、HindⅢ双酶切,酶切产物纯化后连接,连接产物转化大肠杆菌。采用基因特异性引物进行菌液PCR鉴定阳性克隆,重组质粒pSB130-GAFP、pSB130-LTP双酶切产物大小分别为540bp、1200bp,与预期产物大小一致,pSB130-GAFP、pSB130-LTP双T-DNA区双元载体构建成功。载体pSB130-GAFP、pSB130-LTP转化根癌农杆菌后可以直接用于植物的遗传转化。     

外源茉莉酸和真菌激发子诱导白菜CaMBP10的表达

 Recently,CaMBP10 was identified as a plant lipid transfer protein(LTP).In the study,four leaves old cabbages were treated with exogenous jasmonic acid(JA),JA plus LTP,fungal elicitor,200 mmol/L NaCl and 20% PEG respectively for understanding the functions of CaMBP10 in vivo.The expression of CaMBP10 was determined by using semi-quantitative RT-PCR.The results showed that the expressions of CaMBP10 were dramatically up-regulated in different levels under indicated conditions.It demonstrated that CaMBP10 was involved in biotic and abiotic stress response.     

呼和浩特市南郊大气中的氟化物浓度的研究

1990～1991年对呼和浩特市南郊大气中的氟化物浓度用石灰滤纸法进行了测定,对引起某些采样点测定结果偏高的原因进行了分析。讨论了大气氟化物浓度随时间变化的特点。得出了在工作区范围内,呼市南郊是大气氟化物的清洁区的结论。结合1982～1984年农牧学院大气氟化物浓度测定结果,提出在农牧学院附近大气中氟化物浓度呈逐年上升趋势。     

A study of the oxygen plasma treatment on the serviceability of a wool fabric

Low temperature plasma (LTP) treatment using oxygen gas was applied to a wool fabric. The LTP treated wool fabric was tested with several methods: ASTM D5035-1995, ASTM D1424-1996, AATCC Test Method 99-2000, AATCC Test Method 61-2001 1A, AATCC Test Method 15-2002 and AATCC Test Method 8-2001 and the results were compared with the industrial requirements (ASTM D3780-02 and ASTM D4155-01). The results revealed that the LTP treated wool fabric could fulfil the industrial requirements. The results of the investigation were discussed thoroughly in this paper.     

陆地棉LTP家族基因的克隆与表达分析

长链脂肪酸能够通过调节胚珠内源乙烯信号变化来促进棉花的起始和伸长发育,为深入了解棉花脂肪酸的转运过程及参与基因,对棉花全基因组进行了分析,克隆了11个脂质转运蛋白。实时荧光定量PCR证实:LTP6, LTP7, LTP9, LTP11 4个基因在纤维起始时期（野生型与突变体之间）的表达存在差异,这4个基因可能与纤维起始发育有关。通过基因组步移技术克隆了Gh-LTP6基因的启动子,并对其结构进行分析,发现其具有多个MYB结合位点。这些结果为深入研究LTP基因在纤维起始过程中的角色奠定了基础。     

十字花科植物LTP基因BcMF15同源序列克隆与进化分析

为获取LTP在物种间的演化和分类信息,并为验证该基因在花粉发育过程中的生物学功能提供前期证据,根据白菜雄性不育相关的LTP基因BcMF15全长序列设计引物,运用PCR技术分别从十字花科6属10种材料中克隆了LTP基因BcMF15的同源序列。经比较分析表明,这些同源序列的相似性达80%以上,所推导的氨基酸序列相似性达53%以上,且两者种间差异分别为0~1.1%、0~4.0%,除’圆白’萝卜和其他属的差异较大外,属间差异分别是0.1%~23.4%、0.9%~5.0%。LTP基因的氨基酸序列在N端的跨膜疏水结构域高度保守,而在C端则存在一定程度的变异。这些结果表明该基因具有较好的保守性,可能在十字花科植物的花粉发育中行使重要的功能。由核酸序列构建的分子系统树可知,在亲缘进化关系上芸薹属与萝卜属较近,其他依次为荠菜属、拟南芥属和山芥属,而与诸葛菜属最远,可见在十字花科BcMF15序列差异属间较种间大。     

基于二级LTP纹理颜色直方图的动物实时跟踪

目标表征方法是目标跟踪算法当中的关键环节.近来,目标跟踪技术在动物的跟踪以及其他农业领域当中得到广泛应用.但是,只对区域进行一次纹理特征的提取往往不够丰富.因此,对原始提取LTP纹理特征的方法进行改进,提出二级提取LTP特征融合的目标表征方法.通过对原始图像进行相同LTP特征提取,得到目标区域的多级LTP图像特征,将HSV模型中受光照影响较小的H分量作为颜色特征向量,来建立每一级特征相应的直方图.根据嵌入到Mean Shift当中的二级纹理颜色模型来进行动物跟踪.与传统的基于核的跟踪算法进行对比,此方法显著提高动物目标跟踪的鲁棒性.