棉花蔗糖合成酶基因家族在纤维发育期时空表达模式分析

蔗糖合成酶(Sucrose Synthase)是调控植物糖类代谢的关键酶之一,在植物生长发育及渗透调节过程中起着重要的作用。该酶为多基因家族,各成员在植物不同组织及不同发育阶段发挥不同的作用。陆地棉(Gossypium hirsutum)中有15个Sus成员,本研究利用转录组高通量测序及实时定量PCR技术,检测了该基因家族的表达模式。结果表明,Gh Sus1D、Gh Sus3A、Gh Sus3D和Gh Sus6D在起始期(0~3 DPA)大量表达;GhSus7D在10 DPA的纤维中表达量快速增加,达到同期突变体胚珠的4.7倍;Gh Sus1A、Gh Sus5D在纤维伸长期(5~15 DPA)表达量显著高于胚珠,分别在10 DPA、15 DPA时达到峰值;Gh Sus3A、Gh Sus3D、和Gh Sus6D在纤维发育(15 DPA)时表达量重新上升,且在20 DPA时仍处于较高表达水平。Sus基因家族组织表达模式分析表明,它们在陆地棉品种徐州142野生型和突变体中的组织表达模式基本相同,根、茎、叶、花中没有显著差异。但不同Sus基因具有不同的组织表达特异性。     

Cloning and Sequence Analysis of PTTG1 Gene in Luchuan Pig

This experiment was aimed to clone PTTG1 gene CDS sequence of Luchuan pig,and was analyzed by bioinformatics methods.A pair of special primers was designed according to predicted sequence of porcine PTTG1 in GenBank.The coding sequence of PTTG1 in Luchuan pig was amplified by RT-PCR,its gene sequence characteristics and protein structure was systemically analyzed by bioinformatics techniques.The results showed that the cloned PTTG1 fragment included a 609 bp CDS (coding 202 amino acids).The sequence multi-aligned results showed that Luchuan pig shared 90.15%,87.85%,87.52%,87.03%,76.03%,74.38%,55.74% and 44.48% of similar nucleotide sequence with that of Bos,Pan troglodytes,Homos,Macaca,Rattus,Mus,Gallus and Danio retio,respectively.The protein structure analysis results showed that the protein attributed hydrophilic protein without signal peptide,localized in cell cytoplasm and had 16 phosphorylation sites.The phylogentic tree of amino acid indicated that PTTG1 was highly conserved in the process of evolution of different species.The cloning and analysis of PTTG1 gene provided an important foundation for further study biological function of porcine PTTG1 during early embryonic development.     

普通烟草β-胡萝卜素羟化酶2基因克隆与生物信息学分析

β-胡萝卜素羟化酶(beta-carotene hydroxylase,BCH)是植物类胡萝卜素合成代谢中的关键限速酶,为获得烟草β-胡萝卜素羟化酶2(NtBCH2)基因序列,采用同源克隆法从烟草中分离NtBCH2的基因组DNA序列,基因登录号为JX101477。结果表明:1)同源克隆法从烟草中克隆出NtBCH2基因,NtBCH2基因组DNA全长2131bp,cDNA为1 083bp,含7个外显子和6个内含子。2)NtBCH2蛋白有309个氨基酸,分子量为34.79kD,具有7个糖基化位点,11个磷酸化位点,4个跨膜域。3)系统发育树与BLAST分析表明,NtBCH2是番茄的SlBCH和辣椒CaBCH2的同源基因;GENEVESTIGATOR数据库芯片结果表明,NtBCH2在幼嫩的茎和子叶中表达量最高。     

Study on Developmental Expression of ApoCⅡ Gene mRNA in Liver Tissues of Pigs

The aim of this study was to investigate the developmental patterns of ApoCⅡ gene mRNA in liver in Mashen and Large White pigs, and study the relationship between the expression level of ApoCⅡ and the lipid metabolism in pigs.The mRNA relative expressions of ApoCⅡ gene in liver at seven stages of 1,30,60,90,120,150,and 180-day old in Mashen and Large White pigs were determined by quantitative Real-time PCR.The results showed that the developmental trend of ApoCⅡ mRNA expression in liver between Mashen and Large White pigs was different.The ApoCⅡ mRNA abundance was decreased from birth to 60-day old,then increased at 90-day old,and decreased again after that in Mashen pig.However,the relative expression amount in Large White pig was gradually decreased from birth to 150-day old and increased again at 180-day old.Except for the ApoCⅡ mRNA expression amount at 1-day old,the differences of the expression amount at other stages in Mashen and Large White pigs were significant or extremely significant (P<0.05 or P<0.01).The ApoCⅡ mRNA expression in liver was affected by age and breed,and could play an important role in lipid metabolism in pigs.     

Development of IP and SCAR markers linked to the yellow seed color gene in Brassica juncea L.

Previous studies showed that the yellow seed color gene of a yellow mustard was located on the A09 chromosome. In this study, the sequences of the molecular markers linked to the yellow seed color gene were analyzed, the gene was primarily mapped to an interval of 23.304 to 29.402M. Twenty genes and eight markers’ sequences in this region were selected to design the IP and SCAR primers. These primers were used to screen a BC₈S₁ population consisting of 1256 individuals. As a result, five IP and five SCAR markers were successfully developed. IP4 and Y1 were located on either side of the yellow seed color gene at a distance of 0.1 and 0.3 cM, respectively. IP1, IP2 and IP3 derived from Bra036827, Bra036828, Bra036829 separately, co-segregated with the target gene. BLAST analysis indicated that the sequences of newly developed markers showed good collinearity with those of the A09 chromosome, and that the target gene might exist between 27.079 and 27.616M. In light of annotations of the genes in this region, only Bra036828 is associated with flavonoid biosynthesis. This gene has high similarity with the TRANSPARENT TESTA6 gene, Bra036828 was hence identified as being the gene possibly responsible for yellow seed color, in our research.     

Diversity in the carotenoid profiles and the expression of genes related to carotenoid accumulation among citrus genotypes

Carotenoids are not only important to the plants themselves but also are beneficial to human health. Since citrus fruit is a good source of carotenoids for the human diet, it is important to study carotenoid profiles and the accumulation mechanism in citrus fruit. Thus, in the present paper, we describe the diversity in the carotenoid profiles of fruit among citrus genotypes. In regard to carotenoids, such as β-cryptoxanthin, violaxanthin, lycopene, and β-citraurin, the relationship between the carotenoid profile and the expression of carotenoid-biosynthetic genes is discussed. Finally, recent results of quantitative trait locus (QTL) analyses of carotenoid contents and expression levels of carotenoid-biosynthetic genes in citrus fruit are shown.     

欧美杂交杨Pnd-LRR3基因克隆及其抗锈菌侵染表达

为深入探究欧美杂交杨(Populus nigra×P.deltoides)感病的分子机理,以及抗病基因在发病过程中所起的作用,克隆了欧美杂交杨LRR3蛋白编码基因Pnd-LRR3。对Pnd-LRR3基因进行了生物信息学分析,并对其在抗锈菌过程中的功能进行了研究。Q-PCR结果显示,强致病性的落叶松-杨栅锈菌(Melampsora larici-populina)E4菌株接种6 h,Pnd-LRR3基因表达量上调增幅较大,168 h为显著下调。说明Pnd-LRR3基因在E4侵染过程中起到一定抗性作用,但最终无法阻止锈菌的侵染。     

利用钾吸收基因表达评价葡萄叶面喷施钾肥效果和喷施浓度

【目的】叶面喷施钾肥可以快速、 高效地为葡萄补充钾营养,促进葡萄的高产和优质,已经被广泛应用于葡萄生产中。本实验通过对葡萄叶面喷施不同种类及浓度钾肥,测定葡萄叶片和果实等生理指标变化,以及钾吸收相关基因的表达变化,从生理和基因水平上评价这些钾肥的喷施效果,为葡萄生产中钾肥的施用提供一定指导。【方法】本实验以‘夏黑’葡萄为试材,选择两个葡萄生长关键时期盛花期和果实膨大期分别对葡萄叶片喷施0.2%、 0.5%和0.8%三种浓度的K2SO4、 K2CO3、 K2SO4·2MgSO4和KCl。然后对葡萄叶片和新梢的生长率,坐果率,叶绿素含量,单粒重,可溶性固形物等生理指标进行统计分析,并利用荧光定量PCR技术分析4个钾吸收相关基因VvHAK13、 VvKEA2、 VvSIRK和VvSORK的表达情况。【结果】叶面喷施4种钾肥后,葡萄叶片和新梢的生长率,坐果率,叶绿素含量,单粒重,可溶性固形物等各项生理指标均有不同程度的提升。钾肥种类不同,最适喷施浓度不同,同种钾肥在葡萄盛花期和果实膨大期的最适喷施浓度也有所不同,四种钾肥在果实膨大期的最适喷施浓度普遍高于盛花期。四个钾吸收相关基因在喷施不同种类及浓度钾肥后也表现出不同的表达模式,总体来讲VvKEA2、 VvSIRK 、 VvSORK的表达上调,而VvHAK13的表达下调。果实膨大期,需喷施较高浓度的钾肥,钾吸收相关基因才表现出较强烈的响应,而在盛花期则只需喷施较低浓度的钾肥。综合生理指标和基因表达两方面结果,得出4种钾肥效果依次为K2SO4·2MgSO4＞K2SO4＞K2CO3＞KCl。盛花期K2SO4和K2CO3的最适喷施浓度为0.5%,K2SO4·2MgSO4和KCl的最适喷施浓度为0.2%; 果实膨大期K2SO4、 K2SO4·2MgSO4和KCl的最适喷施浓度为0.5%,K2CO3的最适喷施浓度为0.8%。【结论】葡萄叶面喷施钾肥可以有效促进葡萄叶片和果实的生长发育,四种钾肥的效果依此为: K2SO4·2MgSO4＞K2SO4＞K2CO3＞KCl。盛花期K2SO4和K2CO3的最适喷施浓度为0.5%,K2SO4·2MgSO4和KCl的最适喷施浓度为0.2%; 果实膨大期K2SO4、 K2SO4·2MgSO4和KCl的最适喷施浓度为0.5%,K2CO3的最适喷施浓度为0.8%。适宜的喷施浓度可以有效提高钾吸收相关基因的表达,是其提高钾吸收利用的机理之一。     

蛹虫草CmKexin基因克隆和菌丝体中表达检测

为了探讨蛹虫草类枯草杆菌蛋白酶(Subtilisin-like protease)的表达特性,以真菌蛹虫草菌丝体为研究对象,通过RT-PCR获得蛹虫草CmKexin基因的ORF序列,并进行序列分析。应用Northern杂交和Real-time PCR方法,检测蓝光照射后蛹虫草菌丝体中CmKexin基因的转录情况。结果获得蛹虫草CmKexin基因的ORF序列。对翻译蛋白质产物CmKexin进行生物信息学分析,表明该蛋白质含1个属蛋白转化酶的肽酶S8家族功能域(143~429)和1个前体蛋白转化酶P功能域(514~600),符合真菌类枯草杆菌蛋白酶的特征。蛹虫草菌丝体在黑暗预培养4 d后再蓝光照射,Northern Blotting和Real-time PCR检测均可得到相同的结果,即在持续的蓝光照射48~50 h时,出现CmKexin基因的瞬时大量转录。而其他时间内均未检测到该基因的大量转录。试验结果将为蛹虫草类枯草杆菌蛋白酶的利用提供理论依据。     

福建地区2株猪嵴病毒 VP1基因的克隆及遗传进化分析

[目的]调查猪嵴病毒在福建地区腹泻猪群中的流行和变异情况。[方法]根据Gen Bank中登陆的猪嵴病毒(porcine kobuvirus,PKo V)结构蛋白VP1基因序列设计特异性引物,采用RT-PCR方法从某猪场采集腹泻小肠样品中扩增猪嵴病毒VP1基因,将扩增后的目的片段克隆后进行序列测定。应用生物信息学软件,将获得的2株猪嵴病毒VP1和Gen Bank的猪嵴病毒株VP1基因序列进行对比分析。[结果]猪嵴病毒CH/FJNP/12L/2015与匈牙利株K-30-HU/2008/HUN(GQ249161)的核苷酸同源性最高,为88.1%,氨基酸同源性为95.3%。CH/FJNP/12W1/2015与越南株714441/CAOLANH-VH/2012-2-21(KT266058)的核苷酸同源性最高,为88.2%,氨基酸同源性为96.1%,同源重组分析显示,2株毒株均无明显同源重组发生。[结论]频繁的畜禽国际贸易,以及如今便捷的现代化交通工具,人们生活提供方便的同时,也加速了猪嵴病毒毒性的传播。