Light intensity affects the growth and flavonol biosynthesis of Ginkgo (Ginkgo biloba L.)

Response of growth and secondary metabolites to light intensity are useful measurements to determine suitable silviculture treatments for the cultivation of medicinal plants. Here, we analyzed the growth, flavonols (total flavonol, quercetin, kaempferol, and isorhamnetin) content, flavonols yield per plant, and expression of flavonoid biosynthesis-related genes in 2-year Ginkgo (Ginkgo biloba L.) seedlings at four different light intensities (100, 76, 40, and 25 % of full sunlight) in a greenhouse setting. Across all light intensities, the 76 % sunlight treatment produced the highest growth of total biomass, root, stem, and leaf, indicating negative effects of either fulllight or heavy shading on Ginkgo seedling development. Both flavonols (total flavonol, quercetin, kaempferol, and isorhamnetin) content and expression of flavonoid biosynthesis-related genes [PAL (Phenylalanine ammonia-lyase), CHS (Chalcone synthase), F3H (Flavanone 3-hydroxylase), and FLS (Flavonol synthase)] in leaves were highest under 100 % sunlight, suggesting that full sunlight promotes the expression of flavonoid biosynthesis-related genes and increases flavonoid biosynthesis. The highest and lowest flavonol contents were found in leaves and stems, respectively. The 76 % sunlight treatment produced the highest flavonols yield while the 100 % sunlight produced the highest flavonoids content in leaves, indicating that flavonol production per unit land area depends not only flavonol content but also biomass. Overall, in order to achieve the highest flavonols yield per area in Ginkgo leaf-harvesting plantations, it is important to manipulate light conditions of field.     

植物花色素合成的转录调控研究进展

花色素是由植物类黄酮代谢途径产生的次生代谢产物,决定了花、果实和种子的颜色。随着类黄酮代谢途径的研究不断地深入,多种植物编码合成花色素的结构基因也已得到克隆,并研究证明花色素合成结构基因与转录因子(MYB、 bHLH和WD40)有着极其复杂的相互作用模式。本文主要对花青素合成相关的MYB、bHLH和WD40转录因子及其在调节结构基因表达和花青素合成中的作用进行了综述。     

珙桐DiZF-CCCH1基因的克隆及表达分析

【目的】珙桐Davidia involucrata有性生殖能力弱是限制其自然更新的主要原因。为研究珙桐种子发育的分子机制,本课题组前期完成了珙桐种子转录组分析,筛选到1个编号为c37849.graph_c0的转录本,其在正常种子中高量表达,而在发育异常的种子中表达量很低,差异表达显著,因此将其作为研究目标。【方法】使用PCR克隆该转录本的全长序列,并利用在线分析工具对其编码氨基酸的序列特征、蛋白结构和系统进化等进行生物信息学分析预测,采用qRT-PCR检测目标基因在珙桐不同组织部位以及种子不同发育阶段中的表达情况。【结果】序列分析确定目标基因为一个编码CCCH型锌指蛋白转录因子的基因,将其命名为DiZF-CCCH1,该基因开放阅读框全长为711 bp,编码236个氨基酸,无内含子,相对分子量26.74 kDa,为不稳定蛋白,其氨基酸序列含有植物特有的RR-TZF结构域,与来源于甜橙Citrus sinensis的CCCH蛋白同源性最高。基因表达模式分析显示,DiZF-CCCH1在种子发育前期的表达量最高,后期表达量降低,在其他组织中仅有微量表达,推测其可能在种子发育过程中发挥重要作用。【结论】对珙桐DiZF-CCCH1基因进行了克隆与初步生物信息学分析,发现其编码的蛋白属于RR-TZF型锌指蛋白,且DiZF-CCCH1可能是珙桐种子发育过程中的重要调控基因,为进一步探索该基因在珙桐生殖发育过程中的功能奠定基础。     

重金属胁迫下白骨壤数字基因表达谱分析

[目的]从转录组水平研究重金属污水胁迫处理下白骨壤的基因表达变化,以揭示白骨壤响应重金属污染胁迫的机制。[方法]采用基于高通量测序的数字基因表达谱(DGE)技术对重金属污水胁迫处理组和对照组白骨壤材料进行转录组测序分析。[结果]与对照组相比,重金属处理下白骨壤共有10 459个差异表达基因,其中,8 685个表达量上升,1 774个表达量下降。Me V聚类表明:大部分基因在重金属处理样本中的表达量受到极大的诱导。GO功能注释发现,差异表达基因主要定位于叶绿体以及细胞内各种膜结构,参与"细胞代谢"、"细胞组分的组合和生物合成"、"对刺激的响应"等过程。KEGG代谢通路分析显示,差异表达基因分布于126条Pathways,涉及"核糖体"、"光合作用"、"乙醛酸盐代谢"、"丙酮酸代谢"等途径。重金属胁迫还促进萜类、黄酮类化合物生物合成的关键酶基因(牻牛儿基焦磷酸合酶(0009238)、黄酮醇合成酶(0001833))的表达,进而促进白骨壤有效成分的积累;显著诱导细胞分裂素水解酶编码基因CKX7(0057124)、油菜素内脂信号转导组分基因BSK(0043741)等的表达,进而提高白骨壤对重金属胁迫的适应能力。此外,转录因子分析发现,b HLH、NAC、MYB-related和WRKY在重金属胁迫中发挥重要作用。实验选取8个与环境刺激响应密切相关的基因,通过qRT-PCR验证了它们在重金属污水胁迫处理下的表达差异,结果与数字基因表达谱分析的结果较一致,证实了差异表达基因数据的有效性。[结论]重金属处理影响了白骨壤大量的新陈代谢、光合作用、小分子酸合成、激素合成与信号转导及次生代谢产物合成相关基因的表达。     

查尔酮合酶与查尔酮异构酶基因特征及转基因应用

查尔酮合酶和查尔酮异构酶一起构成了黄酮类化合物生物合成的限速酶.底物先是由查尔酮合酶的作用形成袖皮素查尔酮化合物,然后再由查尔酮异构酶异构化成不同的黄酮类化合物.查尔酮合酶和查尔酮异构酶广泛存在于多种植物中,涉及了苯丙氨酸代谢途径中各种具有防御性产物的生物合成,在植物的抗菌机制、抗胁迫、细胞的发育和分化、色素的积累和外源基因的表达等方面起着重要的作用.就查尔酮合酶和查尔酮异构酶的结构特点、编码基因的克隆及序列特征、基因的表达调控以及转基因应用等方面进行了系统的综述,并对其今后进一步的研究提出了适宜的建议.     

转抗菌肽基因毛白杨的培育及其对溃疡病的抗性

以毛白杨为转基因受体材料,利用根癌农杆菌介导法转化来源于益母草的阳离子抗菌肽基因LJAMP2。经卡那霉素筛选,共获得50株抗性植株。GUS组织染色和PCR检测显示有30株抗性植株呈阳性,初步证明外源目的基因已整合到毛白杨基因组中。RT-PCR证实抗菌肽基因LJAMP2在转基因植株中能大量表达。离体抗病性试验表明:转基因毛白杨细胞粗提液的抑菌能力明显强于非转基因植株。进一步将溃疡病菌接种在转基因和野生型毛白杨茎段上培养30天,转基因植株的病级指数均低于非转化植株。上述抗性试验结果表明:在毛白杨中超量表达益母草抗菌肽基因LJAMP2能显著提高其溃疡病抗病性。     

多穗柯4-香豆酸辅酶A连接酶基因的克隆和表达分析

为了克隆多穗柯4-香豆酸辅酶A连接酶(4-coumarate-CoAligase,4CL)基因,了解其表达情况,从而为深入研究4CL基因分子功能及黄酮类化合物生物合成奠定基础,依据多穗柯转录组测序结果,设计特异性引物,PCR扩增得到4CL基因cDNA全长序列,利用相关软件进行生物信息学分析;采用qRT-PCR法检测4CL基因在不同器官中的表达情况,使用SPSS18.0软件分析其表达量与根皮苷含量间的关系。结果表明:多穗柯4CL基因cDNA全长1704bp,包含长1629bp的开放阅读框,编码含542个氨基酸的蛋白质。该蛋白定位于细胞质中,属亲水性蛋白。多穗柯4CL基因在除根以外的各个器官中均有表达,且表达量与根皮苷含量间呈极显著的正相关关系(P<0.01)。     

核桃楸总黄酮及胡桃醌含量测定

研究采取常规提取法、索氏提取法和超声波提取法浸提核桃楸样品(核桃楸的树叶、树皮和外果皮)。采用紫外-可见分光光度法测定核桃楸样品总黄酮含量,高效液相色谱法测定核桃楸样品胡桃醌含量。研究结果表明,总黄酮含量和胡桃醌含量分别为超声波提取法>常规提取法>索氏提取法。3种提取法浸提核桃楸样品,总黄酮含量均有树叶>树皮>外果皮的趋势;胡桃醌含量均有外果皮>树叶>树皮的趋势。超声波法提取核桃楸样品总黄酮含量分别为:树叶5.00%,树皮4.17%,外果皮3.05%;胡桃醌含量分别为:树叶0.002 09%,树皮0.001 31%,外果皮0.002 29%。     

Molecular cloning and expression analysis of a gene encoding a putative beta-ketoacyl-acyl carrier protein (ACP) synthase III (KAS III) from Jatropha curcas

A cDNA clone encoding a putative beta-ketoacyl-acyl carrier protein (ACP) synthase III (KAS III) was isolated from Jatropha curcas L., a woody oil plant. The cDNA clone (named JcKAS III) contained a 1203-bp open reading frame coding for 400 amino acids with a predicted molecular mass of about 42 kDa. The deduced amino acid sequence of the cDNA clone shares about 80% identity to KAS III from other plants, and contains a conserved Cys(176) in the active site and the amino acid motif G(355)NTSAAS(361) which is responsible for binding regulatory acyl-ACPs. Southern blotting analysis indicated that JcKAS III is a single copy gene in the J. curcas genome. Quantitative real-time PCR analysis showed that JcKAS III was expressed in all tissues examined with highest expression in roots, and that expression of JcKAS III increased as seeds developed.     

A new flavone glycoside from the seeds of Tecoma undulata

The new flavonoid 5,7-dihydroxy-6-methoxyflavone 7-O-α-l-rhamnopyranoside (1) was isolated from the seeds of Tecoma undulata.     

越南油茶脂肪酸积累及相关基因表达动态分析

越南油茶(Camellia vietnamensis)茶籽油脂肪酸组成差异是影响其营养和医疗保健价值的重要因素,但目前关于其种子油主要脂肪酸含量动态变化及其分子调控机制仍不清楚。本文以16年生越南油茶实生树种子为试验材料,采用索氏提取法提油,采用气相色谱法鉴定脂肪酸种类和测定脂肪酸含量,采用荧光定量PCR法测定4个关键酶基因的相对表达量。结果表明,越南油茶茶籽油中共含有15种脂肪酸,其中棕榈酸、硬脂酸、油酸、亚油酸、亚麻酸、棕榈油酸和二十四碳一烯酸的含量在其动态变化的不同时期间存在显著变化,且油酸和亚油酸、饱和脂肪酸和不饱和脂肪酸等的含量之间分别呈极显著负相关关系;8月5日到9月5日是油酸快速积累期,且油酸的积累随着种子的成熟而增加,亚油酸的积累则与之相反;不同脂肪酸含量与控制其转化的关键酶基因相对表达量动态变化特点基本一致,关键酶基因的相对表达量与硬脂酸(底物)、亚麻酸和花生一烯酸(最终的生成物)等含量之间具有极显著的典型性相关性;△9硬脂酰-ACP脱饱和酶基因(SAD)相对表达量与硬脂酸含量的偏相关系数显著且呈负相关,硬脂酸含量与脂肪酸脱饱和酶2基因(FAD2)相对表达量、SAD与FAD2和脂肪酸脱饱和酶3基因(FAD3)相对表达量、亚油酸含量与FAD3基因相对表达量等之间的偏相关系数均显著且呈正相关。可见,越南油茶茶籽油存在棕榈酸向硬脂酸、硬脂酸向油酸、油酸向亚油酸和亚油酸向亚麻酸的直接转化过程,选择合适的采果日期是提高茶籽油不饱和脂肪酸比例的有效手段之一,SAD基因表达增强促进硬脂酸向油酸转化,硬脂酸含量对FAD2基因表达、亚油酸含量对FAD3基因表达、SAD基因表达对FAD2和FAD3基因表达等均表现正调控作用。     

Seedlings of Argan (<Emphasis Type="Italic">Argania spinosa</Emphasis>) from different geographical provenances reveal variable morphological growth responses to progressive drought stress under nursery conditions

To alleviate the combined effects of water and heat stress prevailing in drylands, the choice and introduction of appropriate plant species to these conditions is essential for the success of planting in rehabilitation projects. The argan tree (Argania spinosa) is a vigorous plant, admirably adapted to dry climates, with indisputable physiological and ecological characteristics that make of this tree an ideal plant to fight against erosion and desertification process, which seriously threaten arid lands. However, the geographical origin of seeds/seedlings represents a determining factor. In this context, we investigated the morphological responses of growth in two provenances of argan, the provenance of Tindouf ‘PT’ from Algeria and that of Agadir ‘PA’ from Morocco; under water stress conditions. The experiment attempts to evaluate the level of drought tolerance of these two provenances for selecting the planting material that copes and adapts better to hot arid lands. Argan seedlings of both provenances (PA and PT) were submitted to a water stress gradient (75, 40, 20, and 10% of field capacity), then morphological parameters (shoot height, number of leaves, number of spines, root collar diameter, length of taproot) were measured after 3, 6, 9 and 12 months of growth. All variables of morphological growth varied significantly (P < 0.001) between water stress levels, seed geographical provenances and seedling growth ages. The overall of results concerning morphological parameters indicated that the increase of water stress induced in both argan provenances: a decrease in shoot height associated with an increase in length of taproot that resulted in the reduction of leaf numbers and radial growth but the increase the number of spines. The argan plantlets of Agadir have completely failed to tolerate water stress of 10% FC. Under water stress (40, 20, 10% FC), argan seedlings of Tindouf revealed higher growth results than those of Agadir. PT seedlings offset the water deficit by root elongation to ensure growth of the various components of the aerial part. PT seedlings were more resilient to drought stress as compared to those of PA. The highest growth results were obtained with 40% FC in PT seedlings, and with 75% FC among PA seedlings. It is recommended to use the PT seeds in planting projects under drought conditions, while PA seeds are more suitable under conditions of non-water deficit.     

毛竹捕光色素结合蛋白基因结构及表达模式分析

植物通过光合作用将光能转换为化学能,捕光色素结合(LHC)蛋白与色素形成的复合体在捕获、传递和转化光能过程中发挥着重要作用,因此研究毛竹(Phyllostachys edulis)LHC基因结构及表达模式对于揭示其在毛竹光合作用中的功能具有重要意义。采用生物信息学的方法,对毛竹基因组中的LHC基因进行了系统分析。结果表明,在毛竹中共有29个LHC基因同源序列,其包含的内含子数量为0~5个。序列分析表明,29个LHC基因编码的蛋白分别属于光系统Ⅰ和光系统Ⅱ的捕光色素结合蛋白家族LHCⅠ和LHCⅡ。LHCⅠ包含5个亚家族(Lhca 1-Lhca 5),除了Lhca 4含有3个成员外其他亚家族只有1个成员;而LHCⅡ包含6个亚族(Lhcb 1-Lhcb6),每个亚家族的成员不同,其中Lhcb 1的成员最多为7个。亲疏水性预测表明,不同亚家族成员存在着一定差异。蛋白结构预测发现,29个蛋白均包含导肽和成熟蛋白,具有跨膜结构域,均包含色素结合位点;其中12个蛋白的组成以α-螺旋为主,17个蛋白的组成以随机卷曲为主。基因表达谱分析表明,大多数LHC基因主要在叶片和花序中表达,笋中略有表达,而根和鞭中几乎检测不到表达。研究结果为进一步研究毛竹LHC基因的功能奠定了基础。     

Comparative study on the mRNA expression of Pinus massoniana infected by Bursaphelenchus xylophilus

Pine wilt disease(PWD)is a devastating disease affecting the growth of Pinus massoniana,often leading to withering and death.To reveal the changes involved during disease progression,we investigated the mRNA expression profile of P.massoniana infested by Bursaphelenchus xylophilus.The infestation resulted in the downregulation of genes involved in interactions with pathogenic pathways such as disease resistance gene,CC-NBS-LRR resistancelike protein,and the gene encoding a putative nematode resistance protein.Increased infestation pressure(number of nematodes inoculated)caused a continuous decline in the gene expression of stem samples.An infestation of P.massoniana also resulted in a pathway enrichment of genes involved in phenylpropanoid metabolism and flavonoid biosynthesis,which in turn reduced the levels of total phenols and total flavonoids.A downregulation of auxin responsive family protein was observed in infested samples,which resulted in a suppression of plant growth.Thus,upon B.xylophilus infestation,a downregulation of genes associated with the recognition of pathogens,PWD resistance,and growth regulation was observed in P.massoniana,together with a decrease in the levels of phytoalexinlike secondary substances,all of which resulted in withering and ultimately death of P.massoniana.     

‘鲁赫’刺蔷薇籽成分分析及抗氧化、抗炎活性研究

以‘鲁赫’刺蔷薇籽为原料对其营养成分及活性成分含量进行分析,并对提取物进行抗氧化活性和抗炎活性研究。结果表明:‘鲁赫’刺蔷薇籽的营养成分中总糖含量最高,达(24.78±0.65)%,其次是蛋白质,占(15.47±0.16)%。‘鲁赫’刺蔷薇籽中原花青素含量达(12.84±0.65)mg/g,含量较为丰富,总多酚含量次之,为(6.08±0.55)mg/g,总黄酮含量最少,为(1.13±0.11)mg/g。‘鲁赫’刺蔷薇籽具有优异的体外抗氧化能力,对DPPH自由基和羟基自由基的清除率可分别高达(88.87±1.25)%和(89.86±0.90)%。‘鲁赫’刺蔷薇籽对透明质酸酶和白蛋白变性的抑制率分别高达(83.18±0.28)%和(93.29±0.36)%,具有很强的体外抗炎性。‘鲁赫’刺蔷薇籽具有丰富的营养成分和活性物质并具有较强的体外抗氧化、抗炎活性,应用潜力巨大。     

美国栎属种源引种、变异研究:种子及苗期生长变异

对从美国３个州（宾夕法尼亚、纽约和田纳西州）引进的栎属３个树种北方红栎、柳叶栎和红栎的２１个种源进行了两次播种和容器育苗试验。初步掌握了栎属种子砂床播种、芽苗移栽、容器育苗的关键技术。对栎属种子的发芽、芽苗和苗木的生长过程等有了一定的了解。发现栎属不同树种间和种内种源、家系间在种子体积、千粒重、发芽率、３０ｄ萌叶率和苗高生长等性状均存在极显著变异,试验数据表明种子体积北方红栎为大（其中宾夕法尼亚州家系１７２最大）,其次是红栎和柳叶栎。千粒重以北方红栎为重（其中宾夕法尼亚州家系１７２最重）,其次是红栎和柳叶栎。发芽率为北方红栎最高（其中宾夕法尼亚州家系１９１为１００％）,其次是红栎和柳叶栎。３０ｄ萌叶率和苗高生长以柳叶栎为高（田纳西州种源９７０１ＴＮ最高）,其次是红栎和北方红栎。在种间和种内不同种源、家系间种子千粒重与年终苗高生长相关不显著。     

荔枝核活性成分及其提取物抑菌活性分析

为给荔枝副产品的增值利用提供理论依据,对不同荔枝核样品采用香草醛-高氯酸比色法测定了荔枝核总皂苷含量,用硝酸铝-亚硝酸钠比色法测定了总黄酮含量,用蒽酮-硫酸法测定了多糖的含量,分别采用纸碟法和菌丝生长速率法测定了荔枝核提取物对细菌和真菌抑菌圈的大小,并采用混平板稀释法测定了其最小抑菌浓度。结果表明:不同荔枝核样品中总皂苷含量在1.30%～1.40%之间,总黄酮含量在5.78%～6.80%之间,多糖含量在2.85%～3.34%之间。不同品种荔枝核提取物具有不同抑菌活性。妃子笑荔枝核的抑菌活性强于其它品种,其甲醇提取物对细菌的最小抑菌浓度在1～2mg/mL之间。     

油松最佳采种时期的研究

对油松不同采种时期对种子质量影响问题进行了试验研究 ,结果表明 ,各种处理即不同的采种时期对种子质量影响明显 ,从 8月 15日开始 ,到 9月 2 9日的处理 ,种子千粒重、发芽率、发芽势差别极显著。从 8月 15日到 9月 14日种子质量虽有显著差异 ,但都属于不合格种子 ;从 9月 14日到 9月 2 9日种子都达到省标和国标 ,但种子质量随生长期延长 ,种子质量逐渐提高。综合研究可以确定油松最佳采种时期应为 9月 19～ 2 9日     

A new flavone glycoside from the seeds of Shorea robusta

A new flavonoid, 3,7-dihydroxy-8-methoxyflavone 7-O-α- -rhamnopyranosyl-(1→4)-α- -rhamnopyranosyl-(1→6)-β- -glucopyranoside (1), was isolated from the seeds of Shorea robusta.