柑橘黄龙病是一种毁灭性的病害,一旦感染若不能及时发现,就会迅速传播蔓延,殃及整个果园.在尚无有效药剂根治的情况下及时的发现,并铲除病原是十分重要的防止措施.因此,研究开发出精确、快速、方便、易操作的检测方法显得尤为重要.本文针对柑橘黄龙病在田间诊断、血清学检测、分子诊断、光谱检测等方面的快速检测技术进行了综述,并对各种检测技术进行比较分析,以期为未来柑橘黄龙病快速检测技术研究提供参考依据.
前期从香水柠檬自交不亲和花的转录组数据中筛选到一个上调表达的泛素结合酶UBE2I同源基因片段,为了弄清该基因的特性,开展了本研究.以香水柠檬(自交不亲和)和白花柠檬(自交亲和)为试材,根据从香水柠檬花转录组中获得的UBE2I基因片段设计引物,采用同源克隆方法获得了香水柠檬和白花柠檬这2个品种中UBE2I同源基因的ORF全长和DNA全长,对其序列进行生物信息学分析,再对其表达模式进行探讨.结果 显示UBE2I基因在香水柠檬和白花柠檬中的ORF长度均为483 bp,二个品种ORF全长仅有一个核苷酸的差异,编码160个氨基酸,但其氨基酸序列无差异;DNA全长为2 267 bp,拥有有5个外显子和4个内含子,在第2个和第4个内含子序列中存在2个类似于启动子(TATAA)框的结构.UBE2I泛素连接酶蛋白理论等电点为7.64,分子量为17 981.474 44 Da,三级结构中有4个α螺旋、7个β折叠和10个无规卷曲,不是分泌蛋白,没有跨膜结构,肽链呈现疏水性.时空表达分析表明,UBE2I基因在香水柠檬的不同组织中均表达,但在不同组织中的表达存在差异,其中在花粉中的表达量明显高于其它组织,UBE2I在自交(香水柠檬♀×香水柠檬♂)后柱头中的表达量均比(香水柠檬♀×白花柠檬♂)杂交后柱头中的表达量高.本研究为进一步探索发现自交不亲和过程的泛素结合酶UBE2基因奠定基础.
为了探索金柑花发育的分子机制并研究与金柑花发育相关重要基因编码的蛋白质之间的相互作用,本研究以融安金柑不同生长时期的花蕾为材料,采用SMART技术构建了cDNA文库,对该文库进行了鉴定评价.经检测,所构建的cDNA文库滴度为2.83×108 cfu/mL,文库库容为1.42× 1010 cfu,重组率为97.91%,插入的双链cDNA片段长度主要分布在250~2000 bp之间.结果 表明,该文库达到了建库标准,理论上涵盖了全部与融安金柑花蕾发育相关的基因,为后续利用酵母双杂交技术筛选互作蛋白提供了物质基础.
Gametophytic self-incompatibility (GSI) is an important factor affecting fruit development and is one of the important mechanisms of seedlessness in citrus. Although many studies have investigated the mechanisms of SI, this process remains unclear in “Xiangshui” lemon (C. limon). In this study, we cloned one S-RNase homologous gene and one SLF homologous gene from the “Xiangshui” lemon, designated as ClS1 (pistil-specific expression) and ClF-box (pollen-specific expression), which contain open reading frames (ORFs) of 534 and 810 bp, respectively. The expression pattern of these two genes was analyzed by quantitative real-time polymerase chain reaction, and the results showed that the expression pattern of these two genes was significantly upregulated after self-pollination compared to that after cross-pollination. Through pollination in transgenic tobacco, we found that the pollen tubes grew slowly and became twisted after cross-pollination of ClS1♀ and ClF-box ♂ transgenic tobaccos, but the pollen tubes of control, self-pollination ClS1 and cross-pollination ClS1♂, and ClF-box ♀ grew normally. Furthermore, significantly fewer pollen tubes of ClS1♀ and ClF-box ♂ entered the ovules than that in other combinations, and the seed number of cross-pollination ClS1♀ and ClF-box ♂ transgenic tobaccos was significantly reduced. These results indicated that the ClS1 and ClF-box genes contribute to the self-incompatibility of the “Xiangshui” lemon.
College stud ents' education and management is an important element influencing the quality of higher education.Based on the analysis of the punishment of students in a certain college in Guangxi,this paper explores the reform of student punishment system,so as to effectively regulate students' behaviors,maintain students' legal rights,and promote the democracy and legislation of college student management.