橡胶树HbMYB20基因的克隆及其对拟南芥次生壁发育的调控

【目的】MYB转录因子是调控植物木质素合成和次生壁形成的重要转录因子之一。本文分离克隆到一个与拟南芥AtMYB20高度同源的橡胶树MYB转录因子基因HbMYB20,并在拟南芥中对其功能进行研究,以期了解其在橡胶树木质素合成和次生壁发育的分子调控中的作用,为橡胶树木材形成的分子调控机制研究及其遗传改良奠定基础。【方法】采用 blast分析从树皮转录组中筛选出与拟南芥 AtMYB20序列同一性较高的橡胶树 MYB 基因HbMYB20;设计 ORF区特异性引物,以树皮 cDNA 为模板进行扩增得到该目的基因 cDNA 序列。实时荧光定量PCR检测该基因在橡胶树叶片、胶乳、茎干以及木质部与韧皮部的相对表达量。构建 HbMYB20过表达植物载体,使用农杆菌蘸花法转化拟南芥,获得该基因过表达转基因株系。采用乙酰溴法和间苯三酚染色法,分析转基因、野生型拟南芥茎的木质素含量以及木质素在拟南芥茎基部横截面中的分布。对转基因、野生型拟南芥茎基部横截面切片进行甲苯胺蓝染色,并测量分析导管、木质纤维和维管束间纤维细胞的细胞壁厚度。最后,采用实时荧光定量PCR分析转基因及野生型拟南芥木质素和纤维素合成相关酶基因的表达。【结果】克隆得到1个橡胶树 MYB 转录因子基因 HbMYB20,该基因开放阅读框( ORF)为927 bp,编码309aa 的蛋白,氨基酸序列分析显示,HbMYB20与AtMYB20/43和 AtMYB85/42同源性较高,属 R2R3MYB转录因子 G8亚组成员。表达分析显示 HbMYB20在橡胶树茎干和木质部中高表达,胶乳中表达最低。对 HbMYB20过表达拟南芥分析显示,该基因在3个转基因株系中均表达;相对野生型拟南芥,转 HbMYB20拟南芥植株生长抑制,木质部和维管束间纤维的木质素染色面积较少、染色程度变浅,茎的木质素含量和木质纤维、导管及维管束间纤维的细胞壁厚度均显著低于野生型;同时转基因株系中木质素合成关键酶基因4CL1和 CCoAOMT的表达量以及纤维素合成关键酶基因 CesA8的表达显著下调。【结论】橡胶树 MYB转录因子 G8亚组成员 HbMYB20,在茎和木质细胞中高表达。拟南芥中过表达 HbMYB20导致转基因植株的矮小,细胞壁变薄,阻碍木质部中木质素的合成和积累,同时木质素和纤维素合成相关酶基因的表达显著下降。由此推测 HbMYB20对拟南芥的木质素和纤维素合成都具有负调控作用,可能是1个橡胶树次生壁发育的负调控因子。     

胡桃楸天然次生林凋落物分解的生态学过程

采用定时定位采集与监测的方法,对辽东山区胡桃楸天然次生林凋落物分解的生态学过程进行了研究。结果表明:胡桃楸天然次生林凋落物中叶的干物质量比重最大,枝次之,花和果比重较小;凋落物叶中C/N越低,氮元素含量越高,凋落物分解得越快,C/N与凋落物分解速率呈负相关关系;5—9月凋落物干物质平均失重率为17.28%,其中5月失重率最低,为9.73%,随后的6—8月失重率逐月增加,8月失重率最高,为22.37%,9月失重率略有降低,为20.17%;凋落物分解速率与降水量、空气温度、空气湿度、土壤湿度和凋落物自然含水率都呈显著的二次函数关系;影响凋落物分解过程主要环境因子的多元线性回归显著性分析表明,降水量是影响凋落物分解的主导因子,凋落物自然含水率的影响次之。     

蒙古栎次生林林木竞争压力指数研究

为研究林木竞争机制,优化林分结构和调整抚育间伐技术,以汪清林业局蒙古栎次生林为研究对象,构建了蒙古栎自由树的冠幅模型,提出了影响力因子、影响圈共生模式识别因子,量化单木竞争状态。基于108棵蒙古栎优势木数据,采用调整系数法构建蒙古栎自由树的冠幅与胸径相关的冠幅模型为CW=0.109 4+0.299 8×D;采用影响力因子来确定林分中任意对象木的竞争木;将对象木的影响圈与其竞争木的影响圈之间的共生模式划分为4种类型,利用影响圈共生模式识别因子(FSI)进行共生模式判断,分别4种共生模式确定树冠重叠面积的计算方法。基于不同的共生模式,运用相应的竞争压力指数(CSI)计量模型计算CSI值,并通过案例分析,得到样木CSI值为439。     

寄主营养和次生代谢物质对牛角花齿蓟马(缨翅目：蓟马科)产卵选择的影响

为探究牛角花齿蓟马Odontothrips loti的产卵选择及寄主营养和次生代谢物质对其产卵行为的影响,本研究观测了牛角花齿蓟马在苜蓿Medicago sativa、草木樨Melilotus officinalis、红豆草Onobrychis cyri和红三叶Trifolium pratense 4种豆科牧草叶片上的产卵量和卵孵化率,同时测定、分析了牧草主要营养和次生代谢物质含量与蓟马产卵选择的相关性。结果表明,无选择条件下,牛角花齿蓟马在苜蓿上产卵量最高,为59.1粒/复叶,其次是草木樨,15.7粒/复叶,在红豆草和红三叶上的产卵量仅为0.9粒/复叶和1.1粒/复叶。卵孵化率在苜蓿上最高,为85.05%,在草木樨和红豆草上分别为40.09%和39.18%,红三叶上最低,为5.48%。两两选择条件下,牛角花齿蓟马明显偏好在苜蓿叶片上产卵,在其他牧草叶片上不产卵或产卵量极低。相关性分析表明,牛角花齿蓟马的产卵量与寄主可溶性蛋白含量显著正相关(r=0.722,P=0.002),卵孵化率与可溶性蛋白(r=0.673,P=0.004)和淀粉(r=0.586,P=0.017)含量显著正相关。牛...     

长叶榧不同营养器官次生代谢产物含量分析

对长叶榧不同营养器官的鞣质、黄酮、生物碱、游离蒽醌、绿原酸、皂甙、总酚等7种次生代谢产物的含量进行测定和分析。结果显示:长叶榧不同营养器官7种次生代谢产物总量以叶片最高,不同器官的高低顺序依次为叶>幼根>树皮>1年生枝>根>老枝>茎,其中叶的总含量为茎的7.2倍。不同营养器官7种次生代谢产物的含量也以叶片最高,除鞣质含量叶与幼根差异不显著外,其余次生代谢产物含量叶与其它器官的差异均极显著,表明叶可能是次生代谢产物的主要产生器官。幼根的次生代谢产物含量也比较高,幼根也可能合成和积累部分次生代谢产物。     

高粱抗蚜虫性状与次生代谢物关系的研究

通过田间人工接虫、田间调查和室内生化分析研究了不同高粱材料对蚜虫的抗性及其与次生代谢物的关系。结果显示,高粱抗蚜虫性状属单基因显性简单遗传,通过回交转育方式容易获得抗性基因。高粱抗蚜虫性状与植株体内多酚氧化酶活性及丁布（ＤＩＭＢＯＡ）含量间的关系表现出年度间的差异,有时抗性与感性材料间的多酚氧化酶活性和丁布含量都不表现质的不同,说明另有更重要的次生代谢物实际控制着高粱的抗蚜性状。     

湘东丘陵区次生林土壤微生物的分布及酶的活性研究

为了探讨长江中上游低质低效次生林改造途径,摸索综合评价土壤肥力及林分土壤发育状况的方法。研究了次生林不同林分类型土壤微生物数量及酶的活性．结果表明：次生林土壤微生物数量主要集中在0～20cm的土层中,随土层加深,其数量迅速减少;不同林分类型土壤微生物数量相差很大,阔叶林、针阔混交林的数量多而针叶林的数量少;土壤酶的活性以针阔混交林、阔叶林的强．     

Predicting skipjack tuna forage distributions in the equatorial Pacific using a coupled dynamical bio-geochemical model

Skipjack tuna ( Katsuwonus pelamis ) contributes ≈70% of the total tuna catch in the Pacific Ocean. This species occurs in the upper mixed-layer throughout the equatorial region, but the largest catches are taken from the warmpool in the western equatorial Pacific. Analysis of catch and effort data for US purse seine fisheries in the western Pacific has demonstrated that one of the most successful fishing grounds is located in the vicinity of a convergence zone between the warm (>28–29°C) low-salinity water of the warmpool and the cold saline water of equatorial upwelling in the central Pacific (Lehodey et al ., 1997). This zone of convergence, identified by a well-marked salinity front and approximated by the 28.5°C isotherm, oscillates zonally over several thousands of km in correlation with the El Niño–Southern Oscillation. The present study focuses on the prediction of skipjack tuna forage that is expected to be a major factor in explaining the basin-scale distribution of the stock. It could also explain the close relation between displacements of skipjack tuna and the convergence zone on the eastern edge of the warmpool. A simple bio-geochemical model was coupled with a general circulation model, allowing reasonable predictions of new primary production in the equatorial Pacific from mid-1992 to mid-1995. The biological transfer of this production toward tuna forage was simply parameterized according to the food chain length and redistributed by the currents using the circulation model. Tuna forage accumulated in the convergence zone of the horizontal currents, which corresponds to the warmpool/equatorial upwelling boundary. Predicted forage maxima corresponded well with high catch rates.     

Marine Fungal Cerebroside Flavuside B Protects HaCaT Keratinocytes against Staphylococcus aureus Induced Damage

Cerebrosides are glycosylated sphingolipids, and in mammals they contribute to the pro-/anti-inflammatory properties and innate antimicrobial activity of the skin and mucosal surfaces. Staphylococcus aureus infection can develop, not only from minor scratches of the skin, but this pathogen can also actively promote epithelial breach. The effect of cerebroside flavuside B from marine sediment-derived fungus Penicillium islandicum (Aniva Bay, the Sea of Okhotsk) on viability, apoptosis, total caspase activity, and cell cycle in human epidermal keratinocytes HaCaT line co-cultivated with S. aureus, as well as influence of flavuside B on LPS-treated HaCaT cells were studied. Influence of flavuside B on bacterial growth and biofilm formation of S. aureus and its effect on the enzymatic activity of sortase A was also investigated. It was found S. aureus co-cultivated with keratinocytes induces caspase-depended apoptosis and cell death, arrest cell cycle in the G0/G1 phase, and increases in cellular immune inflammation. Cerebroside flavuside B has demonstrated its antimicrobial and anti-inflammatory properties, substantially eliminating all the negative consequences caused by co-cultivation of keratinocytes with S. aureus or bacterial LPS. The dual action of flavuside B may be highly effective in the treatment of bacterial skin lesions and will be studied in the future in in vivo experiments.     

New Ophiobolins from the Deep-Sea Derived Fungus Aspergillus sp. WHU0154 and Their Anti-Inflammatory Effects

Deep-sea fungi have become a new arsenal for the discovery of leading compounds. Here five new ophiobolins 1–5, together with six known analogues 6–11, obtained from a deep-sea derived fungus WHU0154. Their structures were determined by analyses of IR, HR-ESI-MS, and NMR spectra, along with experimental and calculated electronic circular dichroism (ECD) analysis. Pharmacological studies showed that compounds 4 and 6 exhibited obvious inhibitory effects on nitric oxide (NO) production induced by lipopolysaccharide (LPS) in murine macrophage RAW264.7 cells. Mechanical study revealed that compound 6 could inhibit the inducible nitric oxide synthase (iNOS) level in LPS-stimulated RAW264.7 cells. In addition, compounds 6, 9, and 10 could significantly inhibit the expression of cyclooxygenase 2 (COX 2) in LPS-induced RAW264.7 cells. Preliminary structure-activity relationship (SAR) analyses revealed that the aldehyde group at C-21 and the α, β-unsaturated ketone functionality at A ring in ophiobolins were vital for their anti-inflammatory effects. Together, the results demonstrated that ophiobolins, especially for compound 6, exhibited strong anti-inflammatory effects and shed light on the discovery of ophiobolins as new anti-inflammatory agents.