巴西橡胶树乳管细胞HblMYC3互作蛋白筛选与功能分析

MYC是b HLH转录因子家族的亚家族成员,在植物茉莉酸信号转导过程中发挥着重要的调节作用。Hbl MYC3是从巴西橡胶树的乳管细胞中分离鉴定到的MYC类转录因子,其基因表达受割胶和茉莉酸上调。采用酵母双杂交方法初步筛选Hbl MYC3蛋白的互作蛋白,旨在进一步了解Hbl MYC3的功能。结果表明:Hbl MYC1、Hbl MYC2、DNAJ蛋白、谷氧还蛋白2、含A20和AN1锌指结构域的胁迫相关蛋白5、28 ku热和酸稳定的磷蛋白以及25 ku泛素连接酶E2等7种蛋白不同程度地与Hbl MYC3蛋白互作。基于这些互作蛋白的功能,推测Hbl MYC1或Hbl MYC2通过与Hbl MYC3形成二聚体对小橡胶粒子膜蛋白基因表达进行调控,其他蛋白参与胁迫条件下维持二聚体的稳定性和胁迫反应后降解该二聚体。     

TaJAZ7D蛋白在冬小麦JA抗寒途径中的作用

转录抑制因子JAZ（Jasmonate ZIM-domain）蛋白是茉莉酸（Jasmonate,JA）信号转导途径的核心元件,前人研究发现,拟南芥中AtJAZ1、AtJAZ4与AtICE1蛋白互作可抑制 AtICE1基因的转录,负调控拟南芥的抗寒性,然而小麦中TaJAZ与TaICE蛋白的互作调控机制尚不清楚。本研究以强抗寒冬小麦品种东农冬麦1号（Dn1）为试验材料,以与拟南芥AtJAZ1、AtJAZ4蛋白高度同源的TaJAZ7、TaJAZ12蛋白为对象,检测外源茉莉酸甲酯（MeJA）对低温胁迫下 TaJAZ7、 TaJAZ12基因表达量的影响。结果发现, TaJAZ7基因的表达量与温度变化呈明显负相关,故对TaJAZ7(以TaJAZ7D为研究对象进行后续研究)蛋白进行生物信息学分析和亚细胞定位,并利用酵母双杂交技术验证TaJAZ7D蛋白与TaMYC2和TaICE41蛋白的互作。生物信息学分析表明, TaJAZ7D基因编码区序列全长为633 bp,为不稳定的亲水性混合型蛋白;TaJAZ7D蛋白有典型的TIFY和Jas结构域,属于TIFY家族。亚细胞定位发现,TaJAZ7D蛋白位于细胞核中。酵母双杂交验证试验发现,TaJAZ7D蛋白与TaMYC2和TaICE41蛋白分别存在互作关系。     

四株发酵豆乳酵母菌的分离与鉴定

以发酵豆乳为材料,分离出四种酵母菌,并对其进行了初步的分离鏊定和发酵性能试验.结果表明:四种酵母菌在豆乳的发酵过程中,菌体生长代谢旺盛,具有互补性,用其制作的豆乳既有大豆的香味,也有酒精的醇香味,口感易被接受.与乳酸菌发酵豆乳相比,口感有明显的区别.     

小麦转录因子TaMYB5 like转录自激活能力及其参与生理型花粉败育过程表达模式研究

化学杂交剂SQ-1诱导的小麦花粉败育是一个复杂的生理代谢过程,TaMYB5-like被发现参与此过程。为了解TaMYB5-like基因在SQ-1诱导小麦生理型花粉败育过程中的作用,以SQ-1诱导的小麦生理型雄性不育系PHYMS-1376和对照可育系CK-1376四分体、单核早期、单核晚期、二核期和三核期花药为试验材料,克隆TaMYB5-like基因,并对其进行序列结构、生物信息学、表达模式、亚细胞定位及转录自激活效应分析。结果发现,TaMYB5-like基因序列长5 207 bp,其cDNA长为1 133 bp,其中CDS序列长819 bp,编码272个氨基酸,含有2个SANT结构域,分子量为29.36 kDa,无信号肽,无跨膜结构,亚细胞定位在细胞核内;TaMYB5-like基因启动子涉及光响应、逆境胁迫响应、茉莉酸甲酯响应等顺式作用元件。自激活效应发现,TaMYB5-like蛋白的自激活区段位于N端,合适浓度的AbA和3-AT可以抑制BD-TaMYB5-like和BD-TaMYB5-like-N-SANT融合蛋白的自激活性;随着蛋白质氨基酸序列的截断,自激活性呈降低趋势。与CK-1376相比,PHYMS-1376四分体和单核早期花药中TaMYB5-like表达量差异不显著;花药发育到单核晚期、二核期和三核期时,TaMYB5-like表达量显著增加;CK-1376和PHYMS-1376三核期时花药内TaMYB5-like表达量均最高。     

A novel approach for improving yeast viability and baking quality of frozen dough by adding biogenic ice nucleators from Erwinia herbicola

Freezing deteriorates the baking quality of frozen bread dough by causing lethal injury to yeast cells and depolymerization to the gluten network. To investigate the potential of biogenic ice nucleators in frozen food applications, the effect of extracellular ice nucleators (ECINs) from Erwinia herbicola on the baking quality of frozen dough upon three freeze/thaw cycles were investigated. With addition of ECINs to the activity of 2.4 × 10⁶ units per gram of dough, hardening of bread crumb caused by three freeze/thaw cycles was alleviated by about 50% compared to the control. Additionally, the bread from frozen dough with added ECINs showed 50% larger specific volume compared to the control. The mechanism of cryoprotective effects from ECINs was possibly that ECINs helped in preserving the viability of yeast cells during freeze/thaw cycles. ECINs were able to improve the viability of log-phase and stationary-phase yeast cells in suspensions by about 100 and 10 fold, respectively, and viability of yeast in the frozen dough by 17%. This study revealed the potential of ECINs as a cryoprotectant for applications in the food and biotechnology industries.     

利用大豆浓缩蛋白乳清制备水苏糖的发酵条件

利用酵母对大豆浓缩蛋白乳清进行发酵处理制备水苏糖.在确定最佳起始发酵液的糖度为31.8Brix后,对温度、pH、接种量、装液量等工艺条件进行了单因素试验及正交试验,结果表明最佳发酵条件为:温度32°C,pH5.5,接种量8%,在此发酵条件下利用酵母对大豆浓缩蛋白乳清进行发酵处理48 h,水苏糖的纯度达到了90%,保留率为68%.     

Puroindoline-a and puroindoline-b interact with the Saccharomyces cerevisiae plasma membrane through different amino acids present in their tryptophan-rich domain

Puroindolines are two small, basic cysteine-rich proteins isolated from Triticum aestivum seeds and characterized by a tryptophan-rich domain. They form the molecular basis of wheat grain hardness and display antimicrobial activity that may contribute to plant defence. Their antimicrobial activity is presumed to be due to their hydrophobic tryptophan-rich domain. However, little is known about their mode of action and there is no in vivo evidence that the binding of puroindolines to membranes is mediated by their tryptophan-rich domain. In this study, using a yeast complementation assay, we showed that puroindolines interact with the Saccharomyces cerevisiae plasma membrane. By site-directed mutagenesis of their tryptophan-rich domain, we determined that two tryptophan residues (W41 and W44) are mandatory for interaction of puroindoline-a with the yeast membrane whereas interaction of puroindoline-b depends on lysine residues. These results highlight that other residues than tryptophan play a critical role in the interaction of puroindolines with membranes, and probably their affinity for lipids and antimicrobial activities.     

核蛋白筛选系统(NTT)的建立及鉴定

为了研究植物核蛋白的组成及其在各种环境条件下的动态变化,利用核蛋白的核定位信号(nuclear localization signals,NLS)筛选编码核蛋白的基因,建立了一套快速、省力、低成本的核蛋白筛选系统(nuclear transportation trap,NTT).通过鉴定发现,将合成的SV40蛋白大T抗原的核定位信号序列插入筛选载体的多克隆位点,转化酵母EGY48,插入核定位信号的筛选载体转化酵母后可以在选择性培养基(SD/His-/Leu-)上生长,而没有插入核定位信号的筛选载体转化酵母后不能在选择性培养基上生长,从而证明此核蛋白筛选系统有效.比较实验表明,本研究构建的核蛋白筛选系统与已报道的系统相比筛选效率更高.利用此系统从cDNA文库中分离编码核蛋白的基因,对于研究核蛋白的动态组成、了解核蛋白基因在调控细胞发育及其环境胁迫条件下的作用机制具有重要的意义.