甜菜BvGSTU9基因的生物信息学及在镉胁迫下的表达分析

为了进一步研究甜菜谷胱甘肽转移酶BvGSTU9 (LOC104894060)在重金属胁迫过程中的功能。本研究以‘780016B/12优’为实验材料,对该基因序列特征、结构、功能进行预测分析,并利用qPCR检测该基因在不同浓度镉胁迫下的表达量变化。结果显示甜菜BvGSTU9基因全长925 bp,开放阅读框675 bp,编码了由224个氨基酸组成的不稳定膜外蛋白。BvGSTU9与菠菜、藜麦的氨基酸序列相似性较高,与系统发育进化树分析结果基本相符。二级和三级结构预测表明该基因主要由α-螺旋、β-折叠、延伸链及无规则卷曲组成。qPCR显示BvGSTU9基因在不同浓度的镉胁迫下均受到不同程度的诱导,因此可以推断甜菜BvGSTU9基因无论从结构还是功能上,与镉逆境胁迫存在着一定的应答关系。研究结果也为甜菜耐重金属镉机制研究提供参考依据。     

基于GC-MS对水稻种子耐镉差异代谢物的筛选与分析

为了筛选耐镉性较好的水稻品种,本研究利用气相色谱质谱联用仪,对24份不同耐镉程度的水稻品种进行了代谢物的定性与定量分析,通过主成分分析(PCA)和偏最小二乘判别分析(PLS-DA)等多元统计分析方法筛选出水稻耐镉差异性代谢物。结果表明:24份水稻种子共检测出59种主要代谢产物,有机酸类物质可将不同耐镉水平的水稻区分开;同时结合PLS-DA与T检验筛选后得出4种差异代谢物(VIP>1,p<0.05),其中9,12-亚油酸主要存在于低耐品种,而丙二酸、扁桃酸和D-甘露醇主要存在于高耐品种中。说明有机酸类物质与水稻耐镉性相关,可作为区分水稻种子耐镉差异的标志代谢物。     

Antioxidant activity and lipid peroxidation in Artemia nauplii enriched with DHA‐rich oil emulsion and the effect of adding an external antioxidant based on hydroxytyrosol

Artemia nauplii catabolize polyunsaturated fatty acids (PUFA); in particular, they retroconvert docosahexaenoic acid (DHA, 22:6n‐3), so enrichment is a continuous quest towards increasing PUFA through the use of PUFA‐rich enrichment products. However, optimal conditions during enrichment (aeration, illumination and temperatures around 28°C) tend to accelerate autoxidation of PUFA and the formation of potentially toxic oxidation products. Water‐soluble antioxidants like the polyphenolic compound hydroxytyrosol (3,4‐dihydroxyphenylethanol), a polar molecule found in the water fraction resulting after the milling process of olives, arise as promising compounds to prevent oxidation during Artemia enrichments. We investigated the antioxidant activity and lipid peroxidation in Artemia nauplii during enrichment and the effect of adding an external antioxidant based on hydroxytyrosol during the enrichment with a PUFA‐rich emulsion (M70). For this purpose, the activity of antioxidant enzymes (catalase, superoxide dismutase, glutathione‐S‐transferase, glutathione peroxidase), as well as lipid peroxidation, was determined in enriched and unenriched Artemia nauplii. To validate antioxidant activity and lipid peroxidation, in a first experiment, nauplii were enriched with microalgae (Tetraselmis suecica), yeast (Saccharomyces cerevisiae) and M70 emulsion. In a second experiment, enrichment with a commercial emulsion (DC Super Selco), M70, and a combination of M70 and hydroxytyrosol (Hytolive, HYT) added as an external antioxidant were performed. The combination of M70 with HYT produced the best results, in terms of activity of antioxidant enzymes. The analysis of the fatty acids from total lipids showed that the addition of hydroxytyrosol preserved the DHA percentage of enriched nauplii.     

富硒猕猴桃高产栽培技术

鲜嫩的果肉、独特的风味以及丰富的营养价值,使得猕猴桃被称为“维C之王”,微量元素硒对人体的健康至关重要。江苏北部自然资源丰富,适宜种植富硒猕猴桃,且临近上海等沿海经济带,有利于猕猴桃的保质保量运输和及时供应市场。本文主要分析了江苏北部地区优质猕猴桃的配套种植技术,从苗木的选择到高产措施,为高品质富硒猕猴桃的生产提供技术支持。     

镉和模拟酸雨胁迫对苦楝幼苗镉含量及叶片抗氧化系统的影响

为探讨苦楝在重金属Cd和酸雨的单一及复合胁迫下的生理响应特征,采用人工配置土壤盆栽试验,研究不同浓度Cd(0、30、60、90、120、150、180 mg·kg^-1)和3个梯度模拟酸雨(pH值5.6、4.5、3.5)单一及复合处理对苦楝幼苗生物量、Cd含量及叶片保护性酶[超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)]活性和抗坏血酸-谷胱甘肽循环(AsA-GSH)的影响。结果表明,除低浓度Cd(≤30 mg·kg^-1)单一及其与pH值4.5酸雨复合处理对幼苗地上部、地下部生物量有促进作用外,其他处理均表现出抑制作用,其中复合处理对苦楝幼苗的抑制作用更明显,表现为叠加效应;单一Cd处理时,随着Cd浓度的增加,幼苗根、茎、叶中Cd含量逐渐升高,且根系大于茎叶,复合处理抑制了植物对Cd的积累,对叶片的抑制作用最强;Cd和酸雨单一及复合处理下叶片SOD、CAT和POD活性均有所增加,分别在60 mg·kg^-1 Cd、pH值5.6,60 mg·kg^-1 Cd、pH值4.5和90 mg·kg^-1Cd、pH值4.5时出现阈值;在AsA-GSH循环中,单一Cd处理下,AsA、GSH含量和抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)活性均随着Cd浓度的增加呈先上升后下降的趋势,单一酸雨处理对上述4个指标的影响不尽一致,而复合处理下上述指标在Cd浓度≤60 mg·kg^-1、pH值≥4.5时均能保持较高值,且随着胁迫程度的加深均出现了阈值。综上所述,苦楝对重金属Cd和酸雨胁迫具有一定的耐性,可在Cd浓度≤60 mg·kg^-1和pH值≥4.5的酸雨双重污染区域正常生长。本研究结果为土壤Cd污染的酸雨区域的修复植物筛选提供了理论基础和参考依据。     

Effect of organic amendment amount on soil nematode community structure and metabolic footprints in soybean phase of a soybean-maize rotation on Mollisols

It has been well documented that organic amendment affects soil nematode community structure. However, little is known about the effect of organic amendment amount on soil nematodes. To assess the effect of the amount of organic amendments on soil nematode community structure and metabolic activity, the community composition, abundance, and metabolic footprints of soil nematodes were determined in a long-term field experiment with various amounts of organic amendment in Northeast China. Fertilization treatments included an unfertilized control (CK), chemical fertilizer without manure amendment (OM₀), manure applied at 7.5 Mg ha^-1 plus chemical fertilizer (OM₁), and manure applied at 22.5 Mg ha^-1 plus chemical fertilizer (OM₂). A total of 46 nematode genera were found. Treatments with the largest amount of organic amendment had the smallest number of plant parasite genera (5), but a largest number of dominant genera (7). Soil nematodes, bacterivores, and fungivores were the most abundant in OM₂, followed by OM₁, and the lowest in OM₀ and CK. Organic amendment increased the enrichment index (EI), and the large amount of organic amendment increased the metabolic footprints of bacterivore (Baf) and fungivore (Fuf) and enrichment footprint (Ef). The relationships between Baf (or Fuf) and the increases in soil organic carbon (ΔSOC) and total nitrogen (ΔTN) were stronger than those of bacterivore (or fungivore) abundance with ΔSOC and ΔTN, except for the relationship between bacterivore abundance and ΔSOC. The EI and Ef were positively correlated with ΔSOC and ΔTN. These findings suggest that the amount of organic amendment affects soil nematode activity and function at entry levels in soil food web, and that metabolic footprints of soil nematodes may be better indicators than their abundances in assessing their relationships with soil nutrients.     

不同温度下TMV侵染枯斑三生烟的LncRNA差异表达

【目的】 筛选不同温度下烟草花叶病毒（Tobacco mosaic virus,TMV）侵染后枯斑三生烟（Nicotiana tabacum var. Samsun NN）差异表达的长链非编码RNA（long non-coding RNA,lncRNA）,研究lncRNA在枯斑三生烟抗性反应中的作用。【方法】 N基因的温度敏感性使枯斑三生烟在25℃时具备对TMV的抗性、在31℃抗性丧失,在这两个温度条件下对枯斑三生烟接种TMV和磷酸盐缓冲盐水（phosphate buffered saline,PBS）,48 h后提取系统叶总RNA,构建链特异性文库后进行深度测序。对测序结果进行过滤后利用HTSeq将有效数据与近缘品种TN90（N. tabacum var. TN90）基因组比对,筛选得到lncRNA后利用FPKM法估计lncRNA的表达水平。通过edgeR筛选差异表达lncRNA（differentially expressed lncRNA,DElncRNA）,并利用qRT-PCR技术对这一结果进行验证。通过共定位及共表达分析预测DElncRNA的靶基因,通过参考基因组注释、GO和KEGG富集分析研究靶基因的功能。【结果】 4个处理共12个样本经lncRNA-seq各测得约8 000万条clean reads,共获得4 737条已知lncRNA、40 169条新lncRNA。其中64个lncRNA在不同温度条件下TMV侵染后存在差异表达,qRT-PCR测定结果显示这些lncRNA的测序正确率在80%左右,表明本研究所得测序数据具备较高的可信度。对DElncRNA进行靶基因预测,发现一些基因同时被25℃下调和31℃上调的DElncRNA靶向。靶基因注释功能丰富,主要参与植物抗病、激素和代谢等生理过程。部分可能与激素通路相关的lncRNA,在25℃下TMV侵染时呈现下调趋势,而在31℃下TMV侵染则呈现上调趋势。GO富集分析显示靶基因主要参与构成膜、囊泡等组分,具备钙、钾离子通道抑制剂活性等分子功能,使相应离子得以转运引发随后的反应,同时也参与发病、抗原加工和呈现、细胞分裂素代谢等生理过程。KEGG分析发现靶基因显著富集在植物激素信号转导通路,25℃下调和31℃上调的DElncRNA靶基因同时富集在激素信号传导、ABC运输蛋白、苯丙烷类生物合成等通路。【结论】 不同温度（25℃和31℃）条件下TMV侵染枯斑三生烟后,长链非编码RNA差异表达,DElncRNA通过作用于激素信号传导、物质转运等过程参与寄主系统获得性抗性反应。研究结果可为揭示植物系统获得性抗性中lncRNA的调控功能以及新型抗病毒技术开发提供依据。     

以Lorke法为基础的半致死剂量(LD50)分析方法在油酸、化香树果序、黄连素抗镉减毒评价中的应用

镉污染严重着威胁农业的健康发展,如何有效抗镉减毒是保障农业生产安全的重要措施,然而目前缺乏高效的抗镉药物及简便易行的评价方法。本实验借助Lorke法为基础的半致死剂量(LD50)分析方法,检测了油酸、化香树果序、黄连素等不同潜在的抗镉化合物的减毒作用。通过改良后的Lorke法测定不同抗镉药物对小鼠的径口镉LD50的影响。结果显示,与200mg/kg维生素C相比,在灌胃200mg/kg油酸、化香树果序、黄连素能够减轻镉的毒性,提高镉的LD50。对照的LD50由140-160mg/kg分别上升至300-320mg/kg(油酸组),280-300mg/kg(化香树果序组)和320-340mg/kg(黄连素组)。以上结果表明,以Lorke法为基础的LD50分析方法,能够评价出油酸、化香树果序、黄连素的减镉毒作用,及其剂量-效应关系。以上结果同时证明了相同剂量下黄连素效果要优于油酸、优于化香树果序。     

长期秸秆还田和地下水位对土壤镉积累及有效性的影响

以1982年开始的长期定位试验为对象,研究了2个地下水位(高水位-20 cm和低水位-80 cm)下3种施肥方式(高量秸秆还田HS、常量秸秆还田MS、化肥CF)对水稻土镉积累及有效性的影响。结果表明:高水位条件下,长期秸秆还田可增加土壤总镉及有效态镉含量,HS和MS处理分别较CF处理提高21.8%、59.9%和9.8%、49.2%。低水位条件下,HS处理土壤总镉及有效态镉含量高于CF处理,较CF处理分别提高11.2%和31.6%; MS处理土壤总镉及有效态镉含量低于CF处理,较CF处理分别下降8.8%和14.3%。2012年,在保证原定位试验有足够重复的前提下,变更高水位条件下的部分试验处理。当HS处理变更为CF处理5 a后,土壤总镉和有效态镉含量分别较变更前下降2.5%和5.7%;CF处理变更为MS处理后,土壤总镉和有效态镉含量分别较变更前增加16.5%和58.9%。相同施肥处理,高水位土壤总镉含量高于低水位土壤。在MS处理下,高水位土壤有效态镉含量也高于低水位土壤,而在HS和CF处理下,低水位土壤有效态镉含量高于高水位土壤。土壤镉含量和土壤基本理化性质的相关分析表明,土壤总镉与土壤有机质、络合态铁含量极显著正相关,与pH显著负相关;土壤有效态镉与土壤pH、游离氧化铁极显著负相关。研究表明,施肥方式和地下水位是通过改变土壤的基本性质影响土壤镉的积累及其有效形态。     

Heat,wheat and CO2: The relevance of timing and the mode of temperature stress on biomass and yield

Atmospheric CO₂ enrichment affects C3 crops both directly via increased carbon gain and improved water use efficiency and indirectly via higher temperatures and more frequent climatic extremes. Here we investigated the response of spring wheat (Triticum aestivum L. cv. Triso) to CO₂ enrichment (550 vs. 380 µmol/mol) and heat, applied as a constant +4°C increase or a typical heat wave either before or after anthesis, or as two typical heat waves before and after anthesis. We applied a climate chamber approach closely mimicking ambient conditions. CO₂ enrichment increased above‐ground biomass and yield by c. 7 and 10%, but was not able to compensate for adverse heat stress effects, neither before nor after anthesis, with few exceptions only. Yield depression due to heat stress was most severe when two heat waves were applied (?19%). This adverse effect was, however, compensated by CO₂ enrichment. Applying heat stress before or after anthesis did not exert different effects on yield for both +4°C warming and heat wave application. However, +4°C depressed yield more than a heat wave at ambient CO₂, but not so at elevated CO₂. Thus, the interactive effects were complex and prediction of future wheat yield under CO₂ enrichment and climate extremes deserves more attention.