茶树咖啡碱合成酶CRISPR/Cas9基因组编辑载体的构建

CRISPR/Cas9技术是一门新兴的基因组定点编辑技术,具有操作简单、高效的优点,可轻松实现对目标基因的敲除、替换和定点突变等操作。该技术刚诞生,就受到了全球生命科学领域研究者的关注,不到3年的时间就已经成功应用于多种动、植物当中。然而CRISPR/Cas9技术在茶树中的应用面临载体构建问题,本文以茶树咖啡碱合成酶为例,联合采用常规PCR、Overlapping PCR和Golden Gate Cloning技术,构建了包含茶树咖啡碱合成酶双靶点的CRISPR/Cas9基因编辑载体,为CRISPR/Cas9介导的基因组编辑技术在茶树中的应用奠定了坚实基础。     

不同激素对金花茶圈枝繁殖生根的影响

分析NAA、IBA及2种激素等体积混合溶液对金花茶圈枝繁殖生根成活率的影响。结果表明:采用萘乙酸和吲哚丁酸的等体积混合激素溶液1 000~1 500 mg/L能明显提高金花茶圈枝繁殖苗的生根率,达到95%以上。该技术简化了操作程序,可节省人力,在基质方面也节约了生产成本,可直接应用于金花茶苗木大规模生产。     

油茶高接换冠枯桩现象调查与研究

为了探寻影响油茶高接换冠后枯桩的相关因素,为今后油茶高接换冠技术改良提供理论依据,调查研究了嫁接方法、接穗品种、砧木粗度、留截高度和嫁接方位等因素对油茶高接换冠后枯桩现象的影响。结果表明:5种因素对油茶枯桩程度影响极显著。其中,采用插皮腹接、砧木直径2~6 cm枯桩程度最低;采用‘华硕’和‘湘林XLJ14’其枯桩程度显著低于其它品种;留截高度越低枯桩程度越低;嫁接方位在外侧枯桩程度最低。偏相关分析结果表明,留截高度对油茶枯桩程度的影响最大;其次是嫁接方法、砧木直径和嫁接品种;而嫁接方位对油茶枯桩程度的影响最小。     

Changes in peroxidase activity and lignin content of cultured tea cells in response to excess manganese

Abstract

The present study was undertaken to identify the effects of manganese (Mn) on the activity of peroxidase (PO) and the amount of ascorbic acid (AsA) and lignin, as well as the cell viability of suspension-cultured tea cells (Camellia sinensis L. cv. Yabukita). Cells were grown in B5 medium (containing 0.06 mmol L^?1 Mn) and cultured for 24 h in medium with a Mn concentration of 0.9 mmol L^?1 as an excess treatment. No significant difference was observed between cellular growth and the viability of the cultured tea cells after treatment with excess Mn compared with cells grown under control conditions, although the content of Mn in cells in the excess Mn treatment was 12-fold higher than that of the control cells. The amount of total AsA was also not affected by the Mn treatment. The activity of ionically wall-bound peroxidase (IPO) increased in the presence of excess Mn, unlike the content of lignin. Conversely, the activities of soluble PO and covalently wall-bound PO decreased with excess Mn. These findings suggested that IPO might contribute to Mn tolerance in tea cells. However, its role in the mechanism(s) of Mn tolerance has not been elucidated.     

白菜型油菜DFR基因的克隆与序列分析

By using RACE (rapid amplification ofcDNA ends) based homologous cloning strategy, we have successfully isolated the genomic and full-length cDNA sequences of a gene encoding typical DFR (dihydroflavonol-4-reductase) from black-seeded Brassica campestris L. var. oleifera DC.. The gene, designated BcDFR here, is 1 722bp in length and harbors 5 introns with typical splice sites of plant DFR genes. BcDFR cDNA is 1311bp in length with a 1 158bp ORF as well as a 25bp 5‘ UTR and a 128bp 3‘ UTR. The encoded BcDFR protein is 385 aa with a calculated Mw of 42.85kD and a pI value of 5.55. The nucleotide and amino acid sequences of this gene share extensive homologies to plant DFR genes of wide origins especially high similarities to Cruciferous DFR genes. Sequence analyses such as phylogenetic analysis, conserved domain search and substrate specificity region detection all indicated that BcDFR gene is a quite potentially biofunctional gene. Its cloning enables us to further dissect the possible relatedness between DFR gene and Brassica seed coat color traits and to create transgenic novel yellow-seeded rapeseed germplasm through antisense- or RNAi-suppression of DFR gene expression in black-seeded elite cultivars.     

不同激素对碧云茶树丛生芽增殖培养的影响实验

为了获得碧云茶树的组培技术,加快其优良品种繁殖速度,2010～2011年进行了不同浓度的细胞分裂素6-BA和生长素NAA对碧云茶树丛生芽增殖培养的影响。结果表明,以MS为基本培养基,当6-BA为2.0 mg/L,NAA为0.2 mg/L时,对碧云茶树芽的诱导和增殖最有利,增殖率可达2.84;其中NAA差异不显著,6-BA差异显著。     

AM真菌对油茶生长和抗旱性的影响

AM（Arbuscular mycorrhiza）真菌能够促进植物生长，提高植物的抗逆性。AM真菌对油茶生长和抗旱性的影响的研究表明：接种AM真菌促进了油茶生长，改善了油茶叶片水分状况，增强了叶片细胞质膜的稳定性，使可溶性糖含量增加。     

Armillaria species on tea in Kenya identified using isozyme and DNA sequence comparisons

The aim of this study was to identify seven Armillaria isolates obtained from diseased tea bushes in Kenya using pectic enzyme profiles, PCR-RFLP and IGS-I DNA sequence data. The combination of these identification methods confirmed the presence of three distinct Armillaria groups. One of these groups resembled Zimbabwean group I ( A. fuscipes ). The second group was phylogenetically closely related to A. mellea ssp. nipponica . The third group was different from all other African isolates examined, but had isozyme patterns, especially of pectin methylesterases (PMEs), similar to those of isolates related to A. mellea ssp. nipponica. Analyses of sequence data suggested that this group is phylogenetically closely related to A. hinnulea from Australia and New Zealand.     

山茶花红色素的提取及其性质初探

 初步探索了山茶花红色素的提取条件和理化性质, 结果表明, 用料液比1∶10 ( g/mL) 、pH 1(10%盐酸调) 的95%乙醇作提取剂, 在80℃恒温提取6 h, 提取效率较好。山茶花红色素属花色素苷类,pH值对色素影响明显, 在酸性条件下色泽具有热稳定性。光照能加快色素降解, 但与紫外光无关。金属离子Na⁺ 、Ca^{2 +} 、A1^{3 +} 、Cu^{2 +} 、Zn^{2 +}对色泽无影响, Fe^{3 +} 、Pb^{2 +}有不良影响。色素的抗氧化能力较差, 耐还原性较好。蔗糖、葡葡糖和盐等添加剂对色素无影响。     

三个山茶花种(品种) 香气成分初探

 对红山茶品种‘克瑞墨大牡丹’、‘香神’、物种攸县油茶的鲜花香气成分进行了GC /MS分析, 鉴定了‘克瑞墨大牡丹’香气成分37种, ‘香神’香气27种, 攸县油茶34种, 主要以醇、醛、酯、烯、烷及芳樟醇氧化物为主。‘克瑞墨大牡丹’芳樟醇相对含量远高于其它成分, ‘香神’以壬醛和芳樟醇含量最高, 而攸县油茶却以顺- 芳樟醇氧化物Ⅱ和苯乙醇占优势。两个品种在开花前期醇类含量较高, 后期酯类成分含量增加, 攸县油茶在开花前期醇类物质含量高, 越到后期含量越低。