Lipid analysis of three special nervonic acid resources in China

Nervonic acid plays an important role in nutrition and function of the human body. Malania oleifera, Acer truncatum and Xanthoceras sorbifolium are China’s unique woody plant rich in nervonic acid in seed oil. This study aims to investigate the lipid composition of these 3 special resources. Their fatty acids were detected by gas chromatography coupled with flame detector (GC-FID). Triglycerides (TAGs) and phospholipids (PLs) were detected by shotgun-mass spectrometry (shotgun-MS). Results showed that M. oleifera oil presented the highest level of nervonic acid (46.20 ​± ​0.22%) among the 3 oils. Seeds oil of A. truncatum and X. sorbifolium had 3.53 ​± ​0.20% and 1.83 ​± ​0.21% nervonic acid respectively. 53 species of TAGs and 15 species of PLs were identified in M. oleifera oil, with PLs content of 499.94 ​± ​22.34 ​μg/g. In A. truncatum oil, PL and TAG species were twice more than those in M. oleifera oil, and its’ content of PLs was 76.27 ​± ​3.21 ​μg/g. In X. sorbifolium oil, 75 TAGs and 34 ​PLs were detected, with the lowest PLs at 23.84 ​± ​0.17 ​μg/g. The results demonstrated that these 3 vegetable oils have great potential to become nervonic acid supplements for human health.     

Marker gene excision in transgenic Brassica napus via Agrobacterium-mediated Cre/lox transient expression system

Oilseed rape (Brassica napus L.) is one of the most important oil crops in the world. However, study on marker-free transgene of B. napus for bio-safety purpose is limited in this allotetraploid crop. In order to advance marker gene excision research, a newly designed Cre/lox system combining crossing and auto-excision strategy was introduced into B. napus. The system consists of 2 sets of independent vectors including pC35Spro::T7RP carrying T7 RNA polymerase and pCT7pro::Cre carrying T7 promoter respectively. After hybridization of 2 according types of transgenic B. napus, marker gene would be removed as T7 RNA polymerase facilitate T7 promoter to promote Cre gene expression. Totally 52 and 46 positive T₀ transgenic lines of these 2 vectors were obtained after identification by PCR and test trip. T₁ plants from 3 T₀ positive pC35Spro::T7RP lines and 2 T₀ positive pC35Spro::T7RP lines were identified as single copy according to segregation ratio and were chosen for crossing. However, expression of CP4 EPSPS (glyphosate resistance gene) and OXY (bromoxynil resistance gene) were not found in F₁ progeny, which proved that the excision was not complete. The possible reasons for our limited success were investigated and detailed analyses were performed. Although this system is not applicable for generating transgenic B. napus free from selectable marker gene, it provided valuable experience and clue for further improvement of this technique. Many other advantages and further improvement will be progressed in future work.     

甘蓝型油菜对常用稻田除草剂的耐性评价

为降低稻田除草剂残留对油菜生产的风险,筛选不同除草剂耐性的油菜品种,以19个长江流域大面积应用的油菜品种为材料,选择吡嘧磺隆、丙草胺、灭草松、氰氟草酯等4种常用稻田除草剂,通过营养液纸培试验,考查发芽率及幼苗期株高、最大根长、鲜重、干重等性状,以各性状的耐除草剂系数作为衡量指标,对油菜品种进行除草剂的耐性综合评价。结果发现,在稻田除草剂胁迫下,油菜各生长指标反应因基因型而不同,差异显著。通过聚类分析筛选出与除草剂对应的耐性相对较强的油菜品种,并认为主成分分析、隶属函数法和聚类分析的方法综合评价更全面更稳定可靠。在稻-油生产模式中,建议根据水稻除草剂种类,选择对应的耐性油菜品种,降低生产风险。     

甘蓝型油菜MYB28转录因子对根肿菌和水杨酸的响应特征

MYB转录因子参与植物的生长发育和逆境胁迫过程，在植物防御相关信号反应中起着重要的调控作用。 为了明确MYB28在油菜与根肿菌互作中的作用，对感病甘蓝型油菜中双11中MYB28家族基因在根肿菌接种以及 水杨酸处理后的表达模式进行了分析。不同组织表达模式分析显示，相比于根和叶，BnaMYB28 在茎和种子中的表 达水平更高；BnaMYB28 基因在根肿菌和外源水杨酸处理下均有不同程度的表达响应，外源水杨酸处理下Bna⁃ MYB28-1 和BnaMYB28-3 上调表达，BnaMYB28-2 表达则受抑制。根肿菌侵染后，BnaMYB28-1 在整个侵染期表达 量均显著上调，但在加入水杨酸后表达被显著抑制；BnaMYB28-3 表达量在接菌后7 d和28 d显著上调，在关键皮层 侵染期（接菌后14 d）表达量与对照组相比没有变化，但在施加水杨酸处理后表达量显著上调。综上，外源水杨酸能 正向调控BnaMYB28-1 和BnaMYB28-3 表达，BnaMYB28-3 可能通过水杨酸信号路径参与油菜与根肿菌的互作。     

微生物发酵在菜籽饼粕饲用品质改良中的应用研究进展

随着畜牧业的快速发展、优质饲料原料的严重不足以及国际进口贸易形势的日益紧张,开发非常规饲料资源的需求愈发迫切。菜籽饼粕是我国第一大油料作物油菜的加工副产物,饲用价值不高严重制约了在饲料工业中的应用。微生物发酵技术具有改良菜籽饼粕品质和提高畜禽生产性能的优势。本文综述了微生物发酵改良菜籽饼粕饲用价值的机理、影响因素以及微生物发酵菜籽饼粕饲料在畜禽生产中的应用,讨论了目前微生物发酵菜籽饼粕生产中存在的问题,展望了未来微生物发酵菜籽饼粕饲料的发展方向。     

CRISPR/Cas9介导的多基因编辑系统在大豆中的应用

自CRISPR/Cas9（clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9）基因 组编辑技术发现以来，迅速在作物中得到广泛应用。但是，CRISPR/Cas9多基因编辑系统在大豆中的研究尚待开 发。本文利用CRISPR/Cas9介导的多基因编辑系统，分别构建了两个载体，一个载体含6个靶点，编辑7个大豆基因 （4个Glycine max ASYMMETRIC LEAVES1（GmAS1）同源基因和3个GmAS2 同源基因），另一个载体含8个靶点，编辑 11个G. max AGAMOUS 家族同源基因（4个GmAG 同源基因，2个G. max SEEDSTICK（GmSTK）同源基因和5个G. max SHATTERPROOF1（GmSHP1/2）同源基因）。大豆遗传转化后，经表型鉴定和靶点检测发现，CRISPR/Cas9介导的多 基因编辑系统在大豆中成功实现了多基因编辑。当3个GmAS1 同源基因和3个GmAS2 同源基因同时突变时，导致 大豆叶片向远轴面弯曲、皱缩且叶柄变短的表型。当2个GmSHP1 同源基因和2个GmSTK 同源基因同时突变时，导 致豆荚停止发育的不育表型。     

产量和株型性状良好的抗晚斑病花生新种质创制

花生晚斑病抗性常与不良的产量和株型性状相连锁,为发现更多综合性状优良的抗晚斑病品种,以感病亲本中花5号和抗病亲本ICGV 86699及其杂交构建的重组自交系群体（recombined inbreed lines, RIL）为材料,进行晚斑病抗性、产量和株型相关性状的调查,以筛选综合性状优良的抗病新种质。结果表明,4个环境下共发现18个稳定高抗和26个稳定中抗晚斑病的家系;在两个环境中对百果重和单株结果数进行考察,筛选出38个百果重（≥180.0 g）和单株果数（≥20.0个）都比较大的家系;同样,在两个环境中筛选出主茎高（30～60 cm）和总分枝数（≤20.0个）适中的家系54个。综合分析晚斑病病害等级、产量和株型相关性状,共鉴定出4份产量和株型相关性状优良的抗晚斑病新种质,其中1份高抗晚斑病,3份为中抗晚斑病。该研究结果为培育综合性状优良的抗晚斑病花生品种奠定了理论和材料基础。     

Production profile and comparison analysis of main toxin components of Fusarium oxysporum f. sp. sesami isolates with different pathogenicity levels

Fusarium wilt is a common fungal disease in sesame caused by Fusarium oxysporum f. sp. sesami (FOS). To determine the toxin production profiles of the FOS isolates with different pathogenicity levels under various culture conditions, we assessed the content variation of fusaric acid (FA) and 9, 10-dehydrofusaric acid (9, 10-DFA) produced by the four representative FOS isolates. Results indicated that the concentration of FA reached to a maximum of 2848.66 ​μg/mL in Czapek medium, while 9, 10-DFA was mainly produced in Richard and Low-carbon Richard medium. The concentration of 9, 10-DFA on Richard culture medium varied from 0 ​μg/mL to 716.89 ​μg/mL. Of the five culture media used in this study, Czapek culture medium was the most conductive to produce FA. FA production was significantly affected by culture medium, culture time, and their interactions. Results suggest that there is no correlation between toxin production and pathogenicity level of FOS isolates. These findings provide key information for the mechanism analysis of FOS- sesame interaction and pathogen control.     

Determination of the Level and Position of Substitution in Hydroxypropylated Starch by High-Resolution1H-NMR Spectroscopy ofAlpha-limit Dextrins

Eleven hydroxypropylated starch samples were thinned by bacterialalpha-amylase at 80 °C prior to¹H-NMR analysis on a 400 MHz instrument. Hydroxypropyl (HP) levels were calculated from the intensity of the HP methyl signal (1·18 ppm) compared with that of the methylene and methine (HCO) multiplet (3·2–4·1 ppm). HP values determined by¹H-NMR correlated well with those determined by the colorimetric Johnson procedure. Anomeric proton signals in the spectra ofalpha-limit dextrins were used to identify the positions of HP substitution. Methyl 2-O-, 3-O- and 6-O-hydroxypropyl-α-d-glucopyranosides were synthesized, and only the 2-ether shifted the H-1 signal. Eight HP starches with a molar substitution of 0·05–0·23 had 67–78% HP on O-2, 15–29% on O-3 and 2–17% on O–6.