陆地棉转录因子GhNAC7的克隆及功能分析

【目的】从陆地棉中克隆GhNAC7,分析其结构和功能,研究其在棉花不同组织中以及叶片不同发育时期的表达量。并转入拟南芥进一步探究其在棉花叶片衰老过程中的作用。【方法】利用中国农业科学院棉花研究所棉花生物学国家重点实验室建立的棉花衰老叶片cDNA文库中的序列,获得1个含有NAM结构域的EST,使用Oligo6.71设计引物,重新在陆地棉叶片cDNA中进行克隆。使用Gene Structure Display Server软件分析GhNAC7结构,使用在线工具Plant CARE分析启动子序列,利用在线工具Gen Scan进行氨基酸序列翻译。同时,利用拟南芥基因组数据库(TAIR)进行序列比对,选取得分较高的NAC家族基因,使用MEGA 6.06软件和Gene Doc软件进行进化树分析和氨基酸比对。以XbaⅠ和SacⅠ为酶切位点构建35S::GhNAC7-GFP融合表达载体,分析其在洋葱表皮细胞中的瞬时表达,进行亚细胞定位。利用实时荧光定量PCR技术分析GhNAC7在棉花不同组织、不同叶片发育时期以及在200μmol·L~(-1) ABA调控下的表达量。通过构建p GhNAC7-GUS融合表达载体并转拟南芥,分析其启动子特异性。以Eco RⅠ和SalⅠ为酶切位点,利用p BI101和p BI121载体,分别构建融合表达载体并转拟南芥进行过表达分析。【结果】从陆地棉中成功克隆GhNAC7,其全长为1 064 bp,包含3个外显子,2个内含子。生物信息学分析结果表明,GhNAC7开放阅读框为834 bp,可编码277个氨基酸,其蛋白质分子量为31.35 k D,等电点为9.22。结构域分析表明其属于NAC转录因子的NAM亚家族,进化树分析显示GhNAC7与ANAC041、ANAC083同源性最高,其中,GhNAC7与ANAC083结构域位置均为17—58 aa。其启动子核心元件包含一系列与衰老、激素、胁迫相关的顺式作用元件。亚细胞定位表明其蛋白为核蛋白。组织特异性表明GhNAC7在真叶、子叶、花、花药和衰老真叶中均明显表达,其中在衰老的真叶中表达量最高。启动子特异性分析表明,其GUS活性在衰老的叶片中最强。在拟南芥中过表达该基因,转基因植株比野生型表现出明显的衰老症状。荧光定量PCR分析表明,ABA处理后6 h GhNAC7明显上调表达,并在48 h表达量达到最高,这表明ABA可调控GhNAC7表达从而调节棉花叶片衰老。【结论】GhNAC7可以促进棉花叶片衰老并受ABA的调控。     

Nitrous oxide emission from an irrigated cotton field under semiarid subtropical conditions

In a 1-year study, quantification of nitrous oxide (N₂O) emission was made from a flood-irrigated cotton field fertilized with urea at 100kg N ha⁻¹ a⁻¹. Measurements were made during the cotton-growing season (May–November) and the fallow period (December–April). Of the total 95 sampling dates, 77 showed positive N₂O fluxes (range, 0.1 to 33.3g N ha⁻¹ d⁻¹), whereas negative fluxes (i.e., N₂O sink activity) were recorded on 18 occasions (range, −0.1 to −2.2g N ha⁻¹ d⁻¹). Nitrous oxide sink activity was more frequently observed during the growing season (15 out of 57 sampling dates) as compared to the fallow period (3 out of 38 sampling dates). During the growing season, contribution of N₂O to the denitrification gaseous N products was much less (average, 4%) as compared to that during the fallow period (average, 21%). Nitrous oxide emission integrated over the 6-month growing period amounted 324g N ha⁻¹, whereas the corresponding figure for the 6-month fallow period was 648g N ha⁻¹. Subtracting the N₂O sink activity (30.3g N ha⁻¹ and 3.8g N ha⁻¹ during the growing season and fallow period, respectively), the net N₂O emission amounted 938g N ha⁻¹ a⁻¹. Results suggested that high soil moisture and temperature prevailing under flood-irrigated cotton in the Central Punjab region of Pakistan though favor high denitrification rates, but are also conducive to N₂O reduction thus leading to relatively low N₂O emission.     

Mitigation of salinity stress in wheat (Triticum aestivum L.) seedlings through physiological seed enhancements

Salinity stress can be mitigated by using the physiological seed enhancements. Kharchia 65 (salt tolerant) and PI.94341 (salt sensitive) genotypes were evaluated under salt stress (20 dS/m) by various priming treatments, that is hydropriming and halopriming (50?mmol, KCl and NaCl) in a pot study. Experiment was conducted in completely randomized design under factorial arrangements with three replications. Priming agents improved the final emergence percentage (FEP), emergence index (EI) and reduced the mean emergence time of both wheat genotypes under normal and saline conditions as compared to non-primed seed. Priming treatments elicited the efficiency of wheat seedlings by increasing both Fv/Fm and Ft under both normal and salt stress conditions which were strongly linked with low Na⁺ and high K⁺. In conclusion, all three priming agents effectively nullified the deleterious effects of salt stress by improving seed emergence and triggering the physiological attributes of wheat.     

Dicyandiamide application improved nitrogen use efficiency and decreased nitrogen losses in wheat-maize crop rotation in Loess Plateau

Inhibition of nitrification as a mitigation tool to abate nitrogen (N) losses and improve N use efficiency (NUE) is a promising technology. Nitrification inhibitor (dicyandiamide, DCD) was evaluated in two consecutive wheat-maize rotations (2015–2017), with two different N fertilizer levels applied in wheat (160, 220 kg N ha^?1) and maize (180, 280 kg N ha^?1). More NH₄⁺-N contents (101% and 102% in wheat and 74% and 73% in maize) and less NO₃^–-N contents (37% and 43% in wheat and 46% and 57% in maize) were observed at both N levels treated with DCD compared to without DCD. Higher pH, lower EC and reduced NO₃^–-N accumulation were the other benefits of DCD. The NO₃^–-N accumulation within the 0–200 cm soil profile was significantly less at both N levels with DCD (66 mg kg^?1 and 121 mg kg^?1) compared to without DCD (96 mg kg^?1 and 169 mg kg^?1). Application of DCD also improved the growth and yield in both crops. Increase in NUE from 38% to 49% in wheat and 27% to 33% in maize with DCD at higher N level was also observed. Overall, the effectiveness of DCD in retarding the nitrification process was higher in wheat than maize.     

Effect of roots aqueous extract of Delphinium denudatum on morphine-induced tolerance in mice

The aqueous extract of Delphinium denudatum (Dd) roots showed a significant effect against morphine- (10 mg/kg) induced tolerance and dependence in mice. Oral administration of the extract (200-1600 mg/kg) showed a significant dose-dependent inhibition of naloxone- (1 mg/kg) induced withdrawal.     

Epidemiology of gastrointestinal nematodes of sheep managed under traditional husbandry system in Kashmir valley

The present study was conducted with the objective to investigate the seasonal epidemiological prevalence of gastrointestinal tract (GIT) nematodes in different age groups, sexes and breeds (genotypes) of sheep through necropsy and faecal analysis over a period of 2 years in Kashmir valley, India. A total of 1533 sheep were examined [faecal examination: 1035 (year 1: 561, year 2: 474); necropsy: 498 (year 1: 232, year 2: 266)]. Out of these, 945 (61.64%) were found infected [faecal examination: 697 (67.34%, year 1: 390 (69.51%), year 2: 307 (46.99%); necropsy: 248 (49.79%, year 1: 123 (53.01%), year 2: 125 (64.69%)] with GIT nematodes. The over all prevalence of GIT nematodes in sheep in year 1 was 64.76 and 58.37% in year 2 (P=0.04). The parasites in decreasing order of prevalence (%) in sheep were Haemonchus contortus (59.6); Ostertagia circumcincta (38.0); Bunostomum trigonocephalum (37.7); Chabertia ovina (37.7); Trichostrongylus spp. (33.9); Nematodirus spathiger (29.4); Oesophagostomum columbianum (28.4); Trichuris ovis (23.5) and Marshallagia marshalli (22.1). Season, sex, age, and genotype were the factors that influenced the epidemiological prevalence of GIT nematodes in sheep in the present study. The maximum nematode infection was observed in summer season and lowest in winter (P=0.0005). Local Kashmiri breed was less infected as compared to other genotypes (P>0.05). Lower age groups were more infected than adult animals (P>/=0.05). Prevalence was higher in rams (males) than eves (females) (P>0.05). The present study will initially be of great significance to add to the existing knowledge of the epidemiology of GIT nematodes of small ruminants and the findings will be quite helpful to devise the appropriate control and prophylactic strategies for GIT nematodiasis of sheep reared under the temperate agro-climatic conditions.     

A cross-sectional study of methicillin-resistant Staphylococcus aureus at the equine-human interface

Tropical Animal Health and Production - The present study aimed at investigating the percent prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in equines and associated personnel. A...