Isolation of a psychrotolerant Debaryomyces hansenii strain from fermented tea plant (Camellia sinensis) leaves

Abstract

A psychrotolerant, halotolerant and alkalophilic yeast was isolated from fermented leaves of Camellia sinensis Kuntze, the tea plant. The yeast strain, named Tea-Y1, was both phenotypically and genotypically identified as belonging to the species Debaryomyces hansenii. This assignment was confirmed by scanning and transmission electron microscopy. The analysis of growth curves demonstrated the ability this yeast strain to grow in a temperature range between 4°C and 28°C, with an optimum of 23°C. The ecology of this yeast in the C. sinensis phyllosphere, as well as its possible role in tea fermentation and storage, with particular reference to iced tea, are discussed.     

普通油茶叶绿体基因组密码子偏好性分析

为了利用叶绿体基因工程技术改良普通油茶的重要经济性状,该研究以普通油茶叶绿体全基因组序列为材料,从中筛选出51条长度大于300 bp且以ATG起始的非重复CDS(Coding DNA Sequence)为对象,利用CodonW软件分析其密码子偏好性。结果表明:密码子第三位GC含量为27.55%,ENC范围在35.23~56.67之间,平均值为46.09;RSCU值大于1.00的密码子数目为30个,其中29个第三位碱基以U或A结尾;中性绘图表明GC12与GC3的相关系数为0.143,相关性不显著,回归系数为0.0573;频数分布显示,55%基因的ENC比值集中分布在0~0.1,25%基因的ENC比值分布在0.1~0.2之间;对应分析结果表明,第一向量轴占10.12%的差异,第二向量轴占9.36%的差异,其余两轴分别占7.97%和7.46%,前4轴累计差异为34.91%。中性绘图、ENC-plot和对应性分析均表明普通油茶叶绿体基因密码子偏好受突变作用,更多受选择的影响。最终取高表达优越密码子和高频密码子共有的CUU、AUU、GUU、GUA、UAA、CAA、AAA、GAC、GAA、CCU、ACU、GCU、GCA、UGU、CGU、AGU、UUG、GGU等18个密码子作为最优密码子。该研究结果为利用叶绿体基因工程技术改良普通油茶重要经济性状奠定了基础。     

茶树CsMYB123转录因子的克隆及表达特性研究

该实验以茶树品种‘紫娟’为试验材料,利用RT-PCR方法,从茶树cDNA中克隆得到一个R2R3-MYB型基因(CsMYB123)。生物信息学分析显示,CsMYB123基因的开放阅读框为915bp,编码304个氨基酸,蛋白分子量约34.07kD,理论等电点为8.69,含有2个保守的MYB结构域,编码1个R2R3-MYB蛋白;CsMYB123蛋白与拟南芥(Arabidopsis thaliana)MYB转录因子家族第五亚组的AtMYB123亲缘关系最近;CsMYB123属于亲水性蛋白,无N端信号肽,可能定位于细胞核上。荧光定量PCR分析表明,CsMYB123基因在茶树各组织的表达量大小依次为:一芽一叶第二叶第三叶第四叶老茎嫩茎,且在一芽一叶中的表达量是嫩茎的15.68倍;但CsMYB123的表达受IAA、ABA、ETH和GA3的抑制。花青素含量检测显示,茶树‘紫娟’各组织中花青素的含量高低依次为:第二叶一芽一叶第三叶第四叶嫩茎老茎,且第二叶和一芽一叶的含量分别为老茎的15倍和11倍。研究发现,CsMYB123基因在茶树‘紫娟’的新稍中高水平表达,且其表达模式与不同组织中的花青素含量呈较好的正相关关系,推测CsMYB123基因与茶树花青素合成的调控相关。     

茶树CsBAP1基因的克隆与非生物胁迫和激素响应分析

该研究基于茶树转录组数据,从茶树‘龙井43’cDNA中克隆获得茶树CsBAP1基因,对其蛋白质序列特征和基因表达模式以及CsBAP1在茶树不同组织、不同激素处理及不同逆境胁迫下的表达水平进行荧光定量检测。结果显示：（1）克隆得到的茶树CsBAP1基因开放阅读框长度为543 bp,编码180个氨基酸;CsBAP1蛋白为亲水性蛋白,理论相对分子质量为19 398 Da,理论等电点为9.30,含有保守的Ca²⁺依赖性的C2结构域;茶树CsBAP1蛋白二级结构由8.89％的α 螺旋、7.78％的β 转角、35.56％的β 折叠和47.78％的随机卷曲组成;三级结构分析结果显示,CsBAP1包括螺旋和折叠结构,与二级结构吻合。（2）荧光定量分析结果显示, CsBAP1基因在茶树的花、花蕾、幼叶、老叶和成熟叶中均有表达,且在老叶中的表达量最高;外源施用ABA、IAA、MeJA、GA³以及SA均能够抑制茶树CsBAP1基因的表达;在高温（38 ℃）、低温（4 ℃）、干旱（200 g·L^-1 PEG）、高盐（200 mmol·L^-1 NaCl）胁迫下,CsBAP1的表达量均有所上调,且不同时间段的表达存在显著差异。该研究为进一步研究茶树CsBAP1基因的功能奠定了基础。     

Study on biomolecules in extractives of Camellia oleifera fruit shell by GC–MS

This study aims to present an integrated process that can be used to produce biomedical and biological active components from the fruit shell of Camellia oleifera Abel. Through the Foss method, Aldehyde, acid compounds, acyl and alcohol compounds account for 22.7, 15.93, 0.24 and 61.13% of the extractives which were extracted from Camellia oleifera fruit shell by methanol solvents. Furfural, Pyrazole-4-carboxaldehyde, 1-methyl- and 5-Hydroxymethylfurfural account for 4.74, 1.22 and 58.78% of the extractives which were extracted from the fruit shell of Camellia oleifera Abel by ethanol solvents. Aldehyde, acid and amine compounds account for 5.01, 56.18 and 7.20% of the extractives which were extracted from the fruit shell of Camellia oleifera Abel by ethyl acetate solvents. The extractives of fresh flesh of bayberry were rich in rare drug, biomedical and biological activities.     

Diversity and genetic differentiation among populations of Indian and Kenyan tea (Camellia sinensis (L.) O. Kuntze) revealed by AFLP markers

AFLP markers were successfully employed to detect diversity and genetic differentiation among Indian and Kenyan populations of tea (Camellia sinensis (L.) O. Kuntze). Shannon's index of diversity was used to partition the total phenotypic variation into between and within population components. On average, most of the diversity was detected within populations, with 79% of the variation being within and 21% being between populations of Indian and Kenyan tea. A dendrogram constructed on the basis of band sharing distinctly separated the three populations of tea into China type (sinensis), Assam type (assamica) and Cambod type (assamica ssp. lasiocalyx) in a manner consistent with the present taxonomy of tea, the known pedigree of some of the genotypes and their geographical origin. Principal coordinate (PCO) analysis grouped Assam genotypes both from India and Kenya supporting the suggestion that the Kenyan clones have been derived from collections made in this region. The China types were more dispersed on the PCO plot which is a reflection of wider genetic variation. As would be expected, clones collected from the same region exhibited less overall genetic variation. AFLP analysis discriminated all of the tested genotypes from India and Kenya, even those which cannot be distinguished on the basis of morphological and phenotypic traits. Received: 2 May 1996 / Accepted: 14 June 1996     

Lipid degradation during manufacture of black tea

About 85% of the fatty acids liberated during the manufacture of black tea can be attributed to autolysis of 4 major polar lipid classes in tea leaf tissue, phosphatidylcholine, monogalactosyldiglyceride, digalactosyldiglyceride and phosphatidylethanolamine. Linolenic, linoleic and palmitic acids are the principal fatty acids released from these lipids and they all undergo further degradation. Linolenic acid (60% of fatty acids released) is derived mainly from galactolipids and thus the upper limit of release is dependent on the chloroplast maturity and content of the leaf tissues. Lipid breakdown is complete after 2 hr fermentation and, as there appears to be no accumulation of long chain fatty acid intermediates, it is probable that volatile production has also ceased at this time.     

茶梅叶片结构对自然变温的适应

以茶梅叶片为试材,通过田间形态观测、常规石蜡切片和扫描电镜方法,测定茶梅叶片形态、叶片结构和气孔特征在自然变温过程中的适应性变化.结果表明: 随着温度的逐渐降低,茶梅叶色由绿色逐渐变为暗绿色,并出现白色或紫色斑点;相对电导率和枯叶率呈先升高后下降的趋势,3月相对电导率和枯叶率最高,分别为56.0%和25.4%;2014年9月—2015年4月自然变温期间,茶梅叶片中脉厚度、叶片厚度、上下角质层厚度、上下表皮细胞厚度、栅栏组织、海绵组织厚度、中脉突起度、细胞紧实度均呈先上升后下降的趋势,而细胞疏松度先降低后增加,各指标在不同月份间差异显著;2014年10、12月和2015年3月茶梅叶片气孔长度、宽度变化不显著,3月关闭气孔所占比例最高.茶梅通过叶片结构和气孔的关闭提高自身对低温逆境的适应性.