白屈菜CmFAD2基因的克隆及植物转化载体构建

通过对白屈菜低温应答过程的转录组分析发现膜脂不饱和化相关基因的表达在一定过程中发生变化,脂肪酸去饱和酶基因FAD2在随温度的变化趋势为正"V"型,且表达量变化显著。利用NCBI等在线软件对序列进行相关生物学信息分析,并对白屈菜FAD家族成员FAD2基因的完整开放阅读框(ORF)进行克隆,并命名为CmFAD2。选用克隆载体pMD-19-T,转化大肠杆菌DH5α,测序验证序列正确性及完整性。将目的基因与植物表达载体pRI-201-AN连接构建重组DNA pRI-201-AN-Cm FAD2,电击法转化农杆菌LBA4404,利用菌液PCR法验证成功。该基因可作为药用植物抗寒品种创制的候选基因。     

Leaf carbohydrates assimilation and metabolism affect seed yield of rapeseed with different waterlogging tolerance under the interactive effects of nitrogen and waterlogging

Waterlogging is a main factor causing rapeseed (Brassica napus L.) yield loss, and reasonable nitrogen (N) applications can compensate for this loss. To investigate the effects of N rates on seed yield of waterlogged rapeseed, the waterlogging-tolerant rapeseed variety ZS 9 and sensitive variety GH01 were waterlogged for 0 and 10 days with five leaves at the seedling stage under four N rates (0, 90, 180 and 270 kg/ha). Waterlogging significantly decreased seed yield, while N application can alleviate the yield loss. The yield decrease rate of waterlogged GH01 was greater than that of ZS 9 under the same N rate. During the seedling and bolting stage, the leaf photosynthetic rate (Pn) and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity increased, while activities of adenosine diphosphate glucose pyrophosphorylase (AGPase), sucrose synthase (SuSy) and sucrose phosphate synthase (SPS) decreased with more N under the same watering conditions. Compared to the plants without waterlogging, the leaf Pn and Rubisco activity, starch and sucrose contents of waterlogged rapeseed decreased at the two stages; activities of AGPase, SuSy and SPS of waterlogged rapeseed decreased at the seedling while increased at the bolting stage for both the two varieties. At the flowering stage, the Pn, the activities of Rubisco, AGPase, SuSy, SPS and contents of sucrose, starch increased with more N application for both ZS 9 and GH01. Compared to the plants without waterlogging, the Pn and Rubisco activity for waterlogged plants of the two varieties increased; the waterlogged plants of tolerant variety had higher activities of AGPase, SuSy and SPS, while those of sensitive variety was significantly lower. However, the decreased starch and sucrose content were found in both tolerant and sensitive varieties. The activities of AGPase, SuSy and SPS at flowering were highly positively correlated with yield under the interactive effects of N and waterlogging. These results suggested that the flowering stage is the most important stage that N had the positive regulation on waterlogged rapeseed growth. The carbohydrates translocation from leaves to seeds of the tolerant variety were enhanced after waterlogging, while that of the sensitive variety was still inhibited. This was the main reason for the difference in yield between the two waterlogged varieties.     

11个葡萄品种在乐山地区的花芽分化规律

研究了11个葡萄品种3年生树在四川乐山地区避雨栽培条件下的花芽分化表现。结果表明,结果母枝的花芽分化率,魏可、维多利亚、黑芭拉多、金手指、金优2号和巨玫瑰6个品种的都高于50%,较易形成花芽;户太8号、巨峰、夏黑的在43%~48%;美人指和红指较难形成花芽。结果母枝不同节位的花芽分化率,多数品种在第3节位≥50%;结果枝上花穗着生节位在第3.8~4.9节,其中双花率以巨玫瑰、巨峰和户太8号较高,分别为83.45%、73.61%和73.06%。以结果母枝的花芽分化率≥50%节位作为冬剪的最低节位,则金优2号、户太8号、巨玫瑰、金手指、维多利亚5个品种在第3~4节,黑芭拉多和巨峰2个品种在第2~3节,魏可在1~2节。夏黑表现花芽分化率较低,为43.33%,可能与夏黑不耐弱光有关,设施栽培时应加强透光性。     

正交设计优化青蒿SRAP-PCR反应体系及引物筛选

为了建立青蒿的SRAP最佳扩增体系,并筛选出SRAP多态性引物,本研究以青蒿叶片DNA为模板,采用正交试验设计,以Mg^2+、dNTP Mix、Taq DNA聚合酶、引物和DNA模板5种因素5个水平,对青蒿SRAP反应体系进行研究。结果表明,青蒿SRAP-PCR最佳反应体系为:引物0.6μmol/L、Mg^2+2.0 mmol/L、模板DNA 5.1 ng、Taq DNA聚合酶2.0 U、dNTPs 0.25 mmol/L,总体积为25μL。各因素对扩增反应均有不同影响,其中引物浓度的影响最大,dNTPs的影响最小。运用该体系对不同种质资源的青蒿进行验证,证明该体系稳定可靠,并在30个引物组合中筛选出了25对扩增条带清晰,多态性丰富的引物组合。这一结论为今后利用SRAP标记技术进行青蒿分子遗传学研究提供了科学依据。     

同期排卵处理母牛体温和活动量变化规律及效果分析

为探究同期排卵处理母牛体温和活动量变化规律及不同同期排卵技术处理效果，指导同期排卵技术优化。本研究自动监测了18头20月龄左右同期排卵（GnRH-PG-GnRH）处理荷斯坦母牛和17头产后40~60 d预同期排卵（PG-PG-GnRH-PG-GnRH）处理荷斯坦母牛的体温和活动量，应用自动检测系统进行母牛发情监测。结果发现，同期排卵处理母牛发情时阴道温度平均升高（0.43±0.20）℃，持续（12.37±2.73）h；活动量平均升高（18.28±18.61）倍，持续（11.00±1.68）h；排卵时阴道温度平均下降（0.20±0.10）℃，持续（11.00±1.68）h。自动化发情监测显示，同期排卵处理母牛7头发情并排卵；预同期排卵母牛GnRH处理前全部发情排卵。两种同期排卵处理，虽可改变母牛性周期进程，促进母牛性周期同步化，但均难以使母牛性周期完全同步。因此，将同期排卵-定时输精和发情鉴定技术科学结合才能取得更好的繁殖效果。     

番茄褪绿病毒在湖南省首次发生

2016年7月在湖南省蔬菜病害调查中发现,4种茄科蔬菜表现出叶脉间褪绿、叶片黄化等症状,疑似被番茄褪绿病毒(Tomato chlorosis virus,To CV)感染,同时叶片背面聚集了大量烟粉虱。采用To CV热激蛋白(heat shock protein 70,HSP70)序列的特异性引物对采集的番茄、茄子、辣椒、马铃薯样品和烟粉虱样品进行RT-PCR检测,均扩增出目标条带,且扩增序列与北京番茄To CV分离物(KC887999.1)部分序列相似度为99.0%,确认采集样品被To CV感染。4种茄科蔬菜To CV的感染率达70%~100%;发病叶片上烟粉虱的带毒率为66.7%~87.5%;鉴定出烟粉虱的生物类型为MED烟粉虱。这是湖南省首次确认该病毒,需要引起关注和加强防范。     

社会林业与五指山的自然保护

五指山自然保护区是世界上现存地势最高的原始热带雨林之一，是我省目前面积最大的自然保护区，也是海南三大河流的发源地。但由于保护区横跨琼中、通什、保亭三县、市，紧邻保护区还生存着众多贫困的黎、苗族村民。因此，保护区与周边社区的联系是十分紧密的，社区的发展也直接影响到五指山热带天然林的保护。海南省林业局中德海南热带林保护和恢复项目在社会林业与五指山森林的保护上做出了有益的尝试，就在全省来说，也是首次对社会林业与森林保护上进行的探索。五指山地区社会林业的研究，对保护海南热带雨林也有着积极的意义。特对社会林业与五指山的森林保护进行了阐述、分析和设想。     

以球果量、雄花量等数量性状对油松优良无性系的再选择

本研究对油松无性系种子园 2 2个无性系的球果量和雄球花量进行观测调查 ,并对数据进行处理。通过无性系内和无性系间的方差分析、聚类分析和秩序排序法对各无性系进行分类和排序。研究结果表明 :无性系间球果量和雄球花量的差异极显著 ,而无性系内差异不显著。并筛选出 1 61 #,6#,2 9#,63 #,78#5个球果数量大 ,占总数的 46% ,雄花量高 ,占总数的 5 7.6%的无性系为优良无性系     

捕食线虫性真菌菌种保存技术的研究

本试验对捕食线虫性真菌菌种保存技术进行了研究,测定了在不同介质保存条件下,捕食线虫真菌分生孢子和菌丝的存活情况及其不是寄生性线虫的能力.结果表明:在设定的保存条件下,分别经过1周冷冻融化或2周反复冷冻融化,被保存的分生孢子接种于玉米粉琼脂培养基上能生长发育:在加入线虫第3期幼虫后,能生产捕食器,对虫体发挥捕食作用.