Agrobacterium-mediated transformation of the CrDAT gene and selection of transgenic periwinkle lines with a high vincristine accumulation

Catharanthus roseus contains vincristine and vinblastine, which are outstanding drugs for cancer. In the biosynthetic pathways of terpenoid indole alkaloids (TIAs) in C. roseus, deacetylvindoline 4-O-acetyltransferase (DAT) is a key enzyme that catalyses the last reaction of vindoline biosynthesis to form vinblastine and vincristine. In this study, the CrDAT transgene was transferred into the periwinkle by Agrobacterium-mediated transformation and generated transgenic periwinkle lines with an increase in vincristine accumulation. The C. roseus DAT gene was introduced into C. roseus plants and it was confirmed that CrDAT was successfully transferred into the genome of periwinkle plants and efficiently translated to synthesise recombinant DAT protein. Four transgenic periwinkle lines in T1 generation, T1-1, T1-3, T1-6, and T1-7, expressed recombinant DAT protein with the total protein content in the range of 2.86 μg.mg^?1 to 5.12 μg.mg^?1. Moreover, the vincristine contents of four transgenic lines increased by 1.63?2.48-fold compared to non-transgenic plants, ranging from 6.91 µg.g^?1 (fresh weight) to 10.53 µg.g^?1 (fresh weight). The T1-1 line had the highest vincristine content. Hence, the overexpression of the recombinant DAT protein can improve the vincristine accumulation of transgenic C. roseus plants.

Abbreviation: CrDAT - Catharanthus roseus Deacetylvindoline-4-O-Acetyl Transferase; D4H - Deacetoxyvindoline 4-hydroxylase; ELISA - Enzyme-Linked Immunosorbent Assay Monoterpene indole alkaloid; T0, T1 - Generations of transgenic plants; TIAs - Terpenoid indole alkaloids; WT- The wild-type tobacco plants (non transgenic plant); 35S - Cauliflower mosaic virus 35S promoter     

Effects of elevated CO2 concentration on bulbil germination and early seedling growth in Chinese yam under different air temperatures

The present study investigated the effects of elevated carbon dioxide concentration ([CO₂]) and air temperature on the germination of seed bulbils and the seedling vigour of two Chinese yam lines. Plants were grown under two [CO₂] levels, ambient and elevated (ambient + 200 μmol mol^?1), and two mean air temperature regimes, 22.2 °C (ambient + 1.4 °C) and 25.6 °C (ambient + 5.2 °C). Elevated [CO₂] did not affect bulbil germination under both air temperature regimes. During the early growth stage, the dry weight (DW) of leaves, vines, shoots, roots, belowground parts (roots + tubers) and whole plants were higher under elevated [CO₂] than ambient [CO₂] for both lines under the low- and high-temperature regimes. The values of vigour indexes (index I = germination % × seedling length and index II = germination % × seedling DW) were also higher under elevated [CO₂] than ambient [CO₂] for both lines. These results indicated that Chinese yam seedlings respond positively to elevated [CO₂] during the early growth stage. The above:belowground DW ratios were lower under elevated [CO₂] than ambient [CO₂] in seedlings with very small new tubers for both yam lines, indicating that elevated [CO₂] strongly affected the root growth in the early growth stage. The DWs of post-treatment seed bulbils were higher in the elevated [CO₂] under both air temperature regimes. The results showed that Chinese yam used a smaller amount of the reserves in seed bulbils under elevated [CO₂] than under ambient [CO₂].     

中国2种橡胶树炭疽病菌对咪鲜胺敏感性比较

炭疽病是橡胶树上一种重要的叶部病害,主要病原有胶孢炭疽菌和尖孢炭疽菌。为了解中国各植胶区炭疽菌对咪鲜胺的敏感性,采用生长速率法室内测定来自海南、云南、广东和广西4省(区)22株橡胶树胶孢炭疽病菌和15株尖孢炭疽菌对咪鲜胺的敏感性。结果表明:供试的37株橡胶炭疽病菌菌株对咪鲜胺的EC_(50)值为0.022 0~0.272 1 mg/L,平均为(0.061 7±0.055 4)mg/L,EC_(50)最大值是最小值的12.5倍。其中89.2%参试菌株的EC_(50)值介于0~0.100 0 mg/L。根据成组t检验分析结果表明,目前中国4个植胶区的2种橡胶炭疽病菌对咪鲜胺敏感性无明显差异。除了来自海南和云南地区的尖孢炭疽菌有显著性差异外,其他来自不同省份的胶孢炭疽菌和尖孢炭疽菌对咪鲜胺敏感性均无明显差异。研究显示中国橡胶炭疽菌对咪鲜胺仍具有较高的敏感性,但菌株间敏感性有一定的差异,存在一定的潜在抗性风险。     

化学打顶剂在甘肃棉花上的应用效果

2015年在甘肃敦煌植棉区，应用2种化学打顶剂进行不同施用时间和剂量的田间试验表明，土优塔化学打顶剂在6月28日前后、以900 mL·hm^－2的剂量使用，对棉花有较好的封顶效果，对产量及产量构成因子和纤维长度影响不大。2016年示范结果与2015年试验一致     

长沙汉回村民族乡村景观评价研究

采用定量分析与定性分析相结合的理论研究方式,通过景观评价要素筛选和专家打分,确定各个景观要素影响因子的初步权重,并运用网络层次分析法(ANP)分析汉回村作为长沙唯一的少数民族村落景观的合理性,揭示汉回村景观特色主导元素,同时对少数民族与汉族的融合地区乡村景观进行评价,旨在为乡村景观评价提供参考。研究结果表明汉回村乡村景观优势类型:民俗文化景观>自然生态景观>农业产业景观>建筑聚落景观,应保持与发展民俗文化景观优势,改善建筑聚落景观劣势。     

影响北方白菜型冬油菜主要农艺性状变异的气象因子分析

为探究影响北方冬油菜农艺性状的气象因子，以新疆自治区阿勒泰、乌鲁木齐和喀什为研究试点，选择陇油6号、陇油7号、陇油8号、天油2号、天油4号、天油7号共6个抗寒性不同的白菜型冬油菜品种为研究材料，采用适定性分析法和相关性分析法，研究气象因子对冬油菜的株高、主花序角果数、单株角果数、角粒数等农艺性状的影响。结果显示：随纬度降低越冬率增加，农艺性状变劣，其中千粒重、株高和角粒数变异系数较小，分别为5.86%、6.23%和12.63%，单株角果数的变异系数较大，为28.25%；参试品种间，强抗寒品种陇油6号和陇油7号的农艺性状变异系数较小，弱抗寒品种天油2号等变异系数较大；农艺性状与越冬率显著正相关（P<0.05），角粒数与各种气象因子的相关性均达显著水平（P<0.05）；积雪天数、年降雨量和最大积雪厚度对农艺性状具有正效应，累积贡献率达到0.877，最冷月最低气温、地面极端低温、负积温和年均气温对农艺性状具有负效应，累积贡献率达到-0.871。因此，在北方寒区，选择强抗寒冬油菜品种有利于保持农艺性状稳定，获得较好产量与效益。     

鸡大肠杆菌病中草药佐剂多价灭活苗的研制——Ⅰ、疫苗生产工艺

采用鸡大肠杆菌多发病型菌株以及地方病型菌株作为制苗用菌种，保证了疫苗既具有一定的广谱性，又具有一定的特异性；采用液体通气培养方法，在一定程度上提高了菌数；采用中草药蜂胶作为佐剂不但可有效地提高机体的免疫功能，同时兼具有治疗，保健和促生长作用，另外蜂胶还可以增大抗原表面积，延长抗原在组织的贮存时间，可持久地释放抗原，保证了机体持续产生高效价抗体。     

我国南方稻区稗草对二氯喹啉酸的抗药性测定

用琼脂法对2000年和2001年采收的29个稗草生态型对二氯喹啉酸的抗性水平进行了测定。结果表明:广东花都稗草对二氯喹啉酸最为敏感,其抑制中浓度IC50是0.148 0 mg/L;湖南安乡稗草对二氯喹啉酸的抗性极为明显,2000年和2001年样本IC50值分别是7.458和13.80 mg/L,其相对抗性比分别为50.4和93.2;2000年采收的湖南常德稗草也对二氯喹啉酸表现出高抗,其相对抗性比值是4.47,黄梅、汉寿、常德(b)、高桥4地稗草也表现出抗性,其相对抗性比值分别为2.37、3.12、2.44和2.20;其他22个稗草生态型对二氯喹啉酸仍未表现出抗性。     

小麦抗叶锈基因Lr 38的AFLP标记

 小麦叶锈病是小麦的重要病害之一,几乎在所有小麦种植区都有发生,严重时可造成5%~15%甚至更大的产量损失^[1]。小麦抗叶锈基因Lr38发现位于中间偃麦草(Agropyron intermedium)第7组的一条染色体上,并被标定在6 DL上^[2],是抗性很强的抗叶锈病基因,国内外至今尚未发现对Lr38有毒性的菌株,是一个应用潜力很大的抗病基因。     

苦皮藤素类似物的合成与结构鉴定

以苦皮藤Celastrus angulatus Max.提取物水解产物中的多羟基β-二氢沉香呋喃为原料,合成了对粘虫Mythimna separata具有毒杀活性的苦皮藤素(Celangulin)类似物,并在活性追踪的指导下分离得到了两个具有杀虫活性的苦皮藤素类似物 A和B, 其结构经核磁共振谱、快原子轰击质谱、高分辨质谱等波谱学方法鉴定为2β,6α,8β,13-四异丁酰氧基-1β,4α,9α-三羟基-β-二氢沉香呋喃及1β,2β,6α,8β,13-五异丁酰氧基-4α,9α-二羟基-β-二氢沉香呋喃。化合物 A和B 均为新化合物,在20 mg/mL的浓度下对三龄粘虫Mythimna separata的胃毒活性(死亡率)分别为89.5%和93.2%。