Production of Carotenoids and Phospholipids by Thraustochytrium sp. in Batch and Repeated-Batch Culture

The carotenogenic thraustochytrid Thraustochytrium sp. RT2316-16 was grown in batch and repeated-batch cultures using different feeds containing glucose, or glycerol, and yeast extract, for the production of lipids, phospholipids and carotenoids. RT2316-16 produced canthaxanthin, astaxanthin and β-carotene. The effects of biotin, ascorbic acid, light and temperature were evaluated in some of the experiments. In 2-day-old batch cultures, the combined mass percentage of eicosapentaenoic acid and docosahexaenoic acid in total lipids was between 16.5% (glycerol-based medium in the dark; biomass concentration = 4.2 ± 1.1 g L⁻¹) and 42.6% (glucose-based medium under light; biomass concentration = 3.3 ± 0.1 g L⁻¹), decreasing to 3.8% and 6.1%, respectively, after day 4. In repeated-batch cultures, the total lipids in the biomass increased after glucose or glycerol was fed alone, whereas the total carotenoids (168 ± 7 μg g⁻¹ dry weight (DW)) and phospholipids in the biomass increased after feeding with yeast extract. The biomass with the highest content of phospholipids (28.7 ± 4.3 mg g⁻¹ DW) was obtained using a feed medium formulated with glycerol, yeast extract and ascorbic acid. Glycerol was the best carbon source for the production of a biomass enriched with total lipids (467 ± 45 mg g⁻¹ DW). The composition of carotenoids depended strongly on the composition of the feed. Repeated-batch cultures fed with yeast extract contained canthaxanthin as the main carotenoid, whereas in the cultures fed only with glucose, the biomass contained mainly β-carotene.     

多杀菌素微球制备关键工艺研究:Ⅱ

采用乳化-溶剂挥发法,以聚乳酸(PLA)为成球材料(壁材)制备了多杀菌素微球。研究了PLA浓度和油/水相体积比对多杀菌素微球制备的影响规律,确定了制备多杀菌素微球的优选配方及工艺条件。制备得到中位径(D50)为12.73 μm、跨距为1.4811、载药量在31%左右、包封率为100.2%、包封产率为89.4%的多杀菌素微球,重复性良好。扫描电镜(SEM)观察结果表明,所得微球为表面较光滑的实心小圆球。差示扫描量热(DSC)分析结果证实,多杀菌素和PLA的确形成了载药微球。室内毒力测定结果表明,自制5%多杀菌素微球悬浮剂与市售2.5%多杀菌素悬浮 剂(菜喜)对小菜蛾Plutella xylostella 2龄幼虫的毒力基本相同,LC50值分别为0.40和0.38 μg/mL。     

黄腐酸对雾培马铃薯幼苗抗旱性的影响

试验以马铃薯(Solanum tuberosum)幼苗为研究对象,采用雾培装置进行不同黄腐酸用量和干旱胁迫处理(分别为CK,不加黄腐酸和PEG; 10％ PEG;0.01％ FA+ 10％ PEG;0.03％ FA+ 10％ PEG;0.05％ FA+ 10％ PEG和0.10％ FA+ 10％ PEG),分析其对马铃薯生长发育及抗性生理的影响。结果表明,在干旱胁迫下,雾培马铃薯幼苗的成活率、叶绿素含量、膜稳定指数、根系活力、匍匐茎数、结薯率和产量呈下降趋势,而丙二醛(MDA)、过氧化氢(H₂O₂)、脯氨酸、可溶性糖含量和过氧化氢酶(CAT)活性及超氧阴离子(O^-₂)产生速率呈上升趋势,过氧化物歧化酶(SOD)和过氧化物酶(POD)活性先增加后下降,说明雾培马铃薯幼苗在干旱胁迫下生长发育受到显著抑制,正在遭受逆境胁迫。经黄腐酸处理后,幼苗的成活率、匍匐茎数、结薯率和产量则得到了提高,而叶绿素含量、膜稳定指数和根系活力仍呈下降趋势,但较单一干旱胁迫的下降趋势缓慢,而MDA含量、SOD和POD活性及产生速率呈先升高后下降的趋势,脯氨酸、可溶性糖、H₂O₂含量及CAT活性呈上升趋势,其中H₂O₂含量上升趋势较单一干旱胁迫缓慢,说明黄腐酸处理后使干旱胁迫下雾培马铃薯幼苗的生长发育得到了促进,减轻了干旱胁迫对幼苗造成的伤害,提高了植株的整体抗旱性,而这种抗性与黄腐酸浓度呈明显相关性,0.05％ FA对植株的的保护效应最显著,而0.10％ FA则协同PEG加剧了对幼苗的伤害作用。     

水杨酸和乙烯在烟草抗黑胫病中的作用

5mmol/L水杨酸或1g/L乙烯外源处理可诱导烟草对黑胫病的抗性，推迟植株死亡。不能积累水杨酸的转nahG基因植株和不表现乙烯应答反应的转etr1-1基因植株对黑胫病的抗性显著弱于野生型植株。组成性积累高水平水杨酸的转entC和pmsB基因植株对黑胫病的抗性明显强于野生型植株。这些结果说明水杨酸和乙烯增强烟草对黑胫病的抗性，减缓黑胫病的扩展。0．5mmol/L KCN连续浇灌植株4天或浸溃叶片2，4h均不能恢复转nahG基因植株丧失的对黑胫病的抗病性。此结果进一步支持关于KCN处理不能恢复转nahG基因植株丧失的对真菌病害抗病性的假说。     

外源脱落酸对抗寒性狗牙根抗氧化酶活性的影响

以寒敏感的普通狗牙根、海岸狗牙根和抗寒的新农1号狗牙根、喀什狗牙根为供试材料,研究外源脱落酸（0 mg•L^-1、5 mg•L^-1、15 mg•L^-1、25 mg•L^-1）在4 ℃低温处理下（处理时间24 h、72 h）对叶片相对电导率（EC）、丙二醛（MDA）含量、超氧化物歧化酶（SOD）活性、过氧化氢酶（CAT）活性、过氧化物酶（POD）活性的影响。结果表明：与CK2（喷水、常温培养）相比,4 ℃低温处理24 h、72 h,无论是否喷施外源脱落酸,狗牙根均表现为[WTBX]EC[WTBZ]增大,MDA含量增加;但与CK1（喷水、4 ℃低温处理）相比,喷施外源脱落酸可以诱导狗牙根细胞EC、MDA含量降低,促进SOD活性、POD活性、CAT活性的进一步增加,保护膜系统的完整性,增强其对低温的适应。综合分析认为,抗寒狗牙根的适宜外源脱落酸喷施浓度为15 mg•L-1,寒敏感狗牙根为5 mg•L^-1更有利于其对低温的适应。随着低温处理时间的延长,抗寒狗牙根SOD活性增加率、MDA含量降低率明显高于寒敏感狗牙根,说明SOD酶活性不同可能是狗牙根抗寒性差异的原因之一。     

棘孢木霉菌肥对黄瓜枯萎病的防治效果及对连作黄瓜根际土壤微生物种群的影响

为明确棘孢木霉Trichoderma asperellum菌肥在防治黄瓜枯萎病的同时对连作黄瓜根际土壤微生物种群的影响,采用实时荧光定量PCR(real-time fluorescence quantitative PCR,RT-qPCR)和磷脂脂肪酸(phospholipid fatty acid,PLFA)分析方法分别测定了棘孢木霉菌肥对连作4年的黄瓜根际棘孢木霉菌和尖孢镰孢菌Fusarium oxysporum DNA拷贝数变化和黄瓜根际微生物种群的影响。结果表明:棘孢木霉菌肥对黄瓜幼苗期和速长期黄瓜枯萎病的防治效果分别为70.24%和76.81%,均与药剂对照的防效差异不显著。棘孢木霉菌的DNA拷贝数出现2个高峰期,即黄瓜苗期和盛果期,其DNA拷贝数分别为235 000.00 ng/μL和80 500.00 ng/μL。黄瓜速长期、盛果期、生长末期的尖孢镰孢菌DNA拷贝数分别为15.41、54.87和18.36 ng/μL,且显著低于同一时期药剂对照和清水对照。黄瓜幼苗期、速长期、开花期和盛果期根际土壤微生物丰度分别为2.24、1.98、2.52和2.12,均高于药剂对照。由此可见,棘孢木霉菌肥在防治黄瓜枯萎病的同时,还可以改善黄瓜连作土壤微生物种群结构。     

A New Cell Wall Located N-rich Protein is Strongly Induced During the Hypersensitive Response in Glycine Max L.

Soybean (Glycine max (L.) Merill, cv. Williams 82) plants and cell cultures respond to avirulent pathogens with a hypersensitive reaction. After inoculation of soybean with Pseudomonas syringae pv. glycinea, carrying the avirulence gene avrA, or zoospores from the fungus Phytophthora sojae Race 1, a resistance-gene-dependent cell death programme is activated. A new gene was identified by differential display of mRNAs that is specifically activated during the early phase of incompatible pathogen-soybean interactions but does not respond to compatible pathogens. The gene is strongly induced within 2h after addition of P. sojae zoospores. A similar kinetic pattern was observed for P. syringae (avrA) inoculated soybean cell cultures. The gene encodes a deduced protein of 368 amino acids with a very high content of asparagine and was therefore termed N-rich protein (NRP). The protein is composed of two distinct domains, of which only the C-terminal domain has striking homology to proteins of unknown function from other plants. An antibody raised against the recombinant NRP recognizes a protein of 42kDa. The protein is located in the cell wall as indicated by cell fractionation studies. Comparison of the genomic DNA-sequence with the cDNA, identified two introns within the open reading frame. The NRP-gene is not directly induced by salicylic acid or hydrogen peroxide, indicating a distinct and specific signal transduction pathway which is only activated during programmed cell death. The NRP-gene appears to be a new marker in soybean activated early in plant disease resistance.     

12-(取代苯氧异丁酰氧亚氨基)-1,15-十五内酯的合成及除草活性

通过取代苯氧异丁酸与12-(羟基亚氨基)十五内酯的反应,合成了取代苯氧异丁酰氧亚氨基十五内酯,并测定了其对苋菜的除草活性。所有化合物均经过1H NMR、13C NMR和元素分析确证。初步的生物活性测定结果表明,目标化合物 Ⅲa~Ⅲd 的EC50值分别为34.570、46.492、55.385、50.114 mg/L,其活性比对照药剂2,4-D(117.325 mg/L)高,而比苯磺隆(22.381 mg/L)低。     

不同抗逆诱导剂对水稻细胞PBZ 1 基因表达的影响

以水稻品种中花8号的悬浮细胞为试材,分别研究了真菌激发子和盐激发子对用稻瘟灵(IPT)和脱落酸(ABA)预处理24 h的水稻悬浮细胞中PBZ 1 基因表达的影响。结果发现:单独用IPT、ABA处理可以诱导细胞中PBZ 1 基因表达;只用真菌激发子和盐激发子处理的细胞中也可以检测到PBZ 1 基因的表达,但两种激发子对PBZ 1 基因表达的影响不同,在真菌激发子处理后的0~3 h之间PBZ 1 基因表达逐渐增强,而在盐激发子处理后的0~3 h之间则呈现逐渐减弱的趋势,到第3 h就检测不到了;用IPT预处理24 h后,再分别用真菌激发子和盐激发子诱导,水稻细胞中PBZ 1 基因表达强度都显著高于单独用IPT处理的细胞,而且在处理后的第0.5~3 h期间一直保持较强的表达;ABA＋盐激发子处理使细胞中PBZ 1 基因表达推迟且短暂,只在处理后第1~2 h之间可检测到,ABA＋真菌激发子处理使细胞中PBZ 1 基因在0.5~3 h期间一直保持较强的表达,明显强于单独用ABA处理的细胞。可见,IPT和ABA都可以诱导抗病基因PBZ 1 的表达,PBZ 1 基因也可由盐激发子诱导表达,说明PBZ 1 基因对除病害以外的非生物逆境也有积极的作用。