马铃薯AP2/ERF转录因子的克隆及植物表达载体构建

AP2/ERF基因家族转录因子普遍存在于植物中,参与植物体内的各种生物学过程,包括植物的生长发育、生物和非生物胁迫响应等。前期转录组测序的结果发现马铃薯‘加湘1号’一个AP2/ERF家族基因(PGSC0003DMG400012154)在接种晚疫病菌(Phytophthora infestans)24 h后被显著激活。从接种P.infestans 24 h的‘加湘1号’的总RNA中通过RT-PCR获得了该基因CDS序列为885 bp,BLAST分析显示该基因编码一个295个氨基酸残基的蛋白,并且含有1个AP2/ERF结构域,是AP2/ERF转录因子家族中的ERF亚家族的一员。本研究将该基因与XcmⅠ酶切的表达载体pCXSN连接,转化大肠杆菌,通过测序挑选插入正确克隆酶切验证,并成功转化农杆菌。本研究结果为进一步研究该基因的功能提供了帮助。     

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。     

中国竹类多样性及其重要价值

竹类属于禾本科Gramineae（Poaceae）、竹亚科Bambusoideae植物，全世界约1 500多种。中国竹类的自然分布广阔，横跨热带、亚热带和温带地区，涵盖包括台湾、香港在内的21个有竹子分布的省、自治区、直辖市和特区。中国是世界上竹类多样性最丰富的国家，也是世界上栽培竹类最丰富的国家，共有竹类植物753种（含22种引进种），约占世界总数的50%以上。同时，中国竹类在起源、遗传进化、内部构造、生态及林型等方面均具有十分丰富的多样性。中国竹类从宏观到微观所展现出的多样性特质，是大自然留给人类的宝贵遗产，对于人类认识自然、保护自然、从而更好的利用竹类资源服务于人类的长远利益，具有重要价值。文章从起源、生态、林型、物种、形态、遗传等方面概述了中国竹类的多样性，为进一步探讨竹类多样性提供基础信息。     

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。     

狗头枣GSTU基因的克隆及其序列分析

为探索红枣GSTU类基因在红枣抗逆中的分子作用机制,本研究以狗头枣枣树叶片的DNA为模板,根据枣GSTU基因的序列(HM345954.1)设计引物,采用PCR方法获得GSTU基因序列,并利用生物信息学手段对其所对应蛋白的结构域、功能域、理化特性、跨膜区、二级结构及亚细胞定位等进行分析。结果表明:克隆到的DNA序列全长838 bp,具有2个外显子和1个内含子,内含子在292~460 bp碱基处,该序列与GenBank中枣的GSTU核苷酸序列和氨基酸序列的同源性分别为99.85%和100%,表明成功的克隆了狗头枣的GSTU基因。该序列编码222个氨基酸,分子量为25.268 kD,理论等电点(pI)为6.10;亲水性的平均数为-0.242,表明该蛋白是亲水性蛋白。二级结构主要以α-螺旋为主,其次是无规则卷曲,不存在信号肽,表明该蛋白可能为非分泌性蛋白,该蛋白可能位于内质网(膜)和质膜上。本研究获得的狗头枣GSTU基因在基因结构上含有1个内含子,在核酸序列及氨基酸序列上与GenBank中枣的GSTU基因同源性高,并对其理化性质、结构等进行了预测与分析,为进一步研究红枣抗逆分子机制提供理论基础,同时为红枣品种的选育提供理论支持。     

Effect of nitrogen root zone fertilization on rice yield,uptake and utilization of macronutrient in lower reaches of Yangtze River,China

Improper application of nitrogen (N) has led to high N losses and low N use efficiency in the lower reaches of Yangtze River in China. An effective method to solve such problems is the deep fertilized N in root zone (RZF). Limited information is available on the effect of RZF on the uptake of macronutrients (N, P and K) and rice yield. Field experiments, conducted from 2014 to 2015, compared the farmer fertilizer practice (FFP, with 225 kg ha^?1 of N, split into three doses) and RZF using the same rate but placing N 5 cm away from rice roots in holes 10 cm deep (RZF10) or 5 cm deep (RZF5) as a single application. The highest mean yield (10.0 t ha^?1) was obtained in RZF10, which was 19.5% more than that in FFP. Root zone fertilization of urea (whether 10 cm deep or 5 cm deep) resulted in greater accumulation of N, P and K in stem, leaf sheaths, leaf blades and grains compared to that in FFP in sandy and in loam soils. The uptake of N, P and K was the highest in RZF10 (average at 176.7, 66.2 and 179.1 kg ha^?1, respectively), higher than that in FFP by 45.0, 17.0 and 22.6%, respectively. N apparent recovery efficiency was markedly higher in RZF10 (53.1%) than in FFP (27.5%). RZF10 significantly increased the N, P, K uptake compared with FFP under different N rates in both sandy and loam soils. These results suggest that the N, P and K input amount should be re-determined under RZF.     

氟铃脲在韭菜中的残留分析及膳食风险评估

为评价氟铃脲在韭菜中使用的安全性,开展氟铃脲在韭菜中的残留量及残留降解研究。进行1年4地田间试验。消解动态试验按氟铃脲11250 g/ha(562.5 g a.i/ha,推荐最高剂量的1.5倍)施药;最终残留试验按氟铃脲11250 g/ha(562.5 g a.i/ha,推荐最高剂量的1.5倍)和7500 g/ha(375 g a.i/ha,推荐最高剂量)施药,施药1~2次,施药间隔7d,1次施药按照药后间隔10、14、21d采集韭菜样品;2次施药按照末次药后间隔7、10、14d采集韭菜样品。液相色谱串联质谱法对氟铃脲进行定量分析。田间消解动态试验表明:氟铃脲在韭菜中消解较快,在山东保护地和安徽露地半衰期分别为7.7和11.5d。膳食风险评估结果表明,氟铃脲在韭菜中的残留风险处于安全水平。建议使用5%氟铃脲乳油防治韭蛆,用药量7500~11250 g/ha,韭菜收割后2~3d,根部喷淋,最多施药2次,施药间隔7d,安全间隔期为14 d。