翠竹及菲白竹LEA3同源基因的克隆及序列分析

竹类植物在中国有着非常悠久的栽培历史,并被广泛应用于园林、造林、庭院以及食品等行业。本研究以翠竹和菲白竹为材料提取其叶片总DNA,采用PCR方法获得翠竹SpLEA3基因与菲白竹SfLEA3-1、SfLEA3-2基因;其中SpLEA3全长804 bp,编码195个氨基酸,GC含量为68.4%;SfLEA3-1全长803 bp,编码195个氨基酸,GC含量为68.2%;SfLEA3-2全长557 bp,编码144个氨基酸,GC含量为67.8%。通过ProtParamy等生物软件分析,SpLEA3、SfLEA3-1和SfLEA3-2与贵州悬竹LEA3基因同源性高达61%以上,且3个基因编码蛋白均属于亲水性蛋白。源自2个竹种LEA3基因均包含一个完整的开放阅读框,其编码的氨基酸含有4~6个由11个氨基酸组成的保守基元序列。本研究不仅为深入了解竹类植物抗旱的分子机理研究提供了基础数据,也为竹类植物的抗旱育种后续研究提供了科学依据。     

碱胁迫下榆叶梅差异基因表达分析

为了解榆叶梅(Prumus triloba Lindl.)在短期碱胁迫下的分子响应机制,本研究采用Illumina HiSeq^TM2500高通量测序技术对其在0、1、3、6、12 h碱胁迫下的叶片进行了de novo转录组测序,以此鉴定与碱胁迫相关的差异表达基因(DEGs)。结果表明,从5 960万条raw reads中获得了约5 300万条高质量clean reads,经从头组装后获得了124 786条榆叶梅unigenes。基于8 948个 unigenes差异表达(DEGs), GO富集分析发现,其中有28个基因可能在早期碱胁迫响应机制中起着重要作用; KEGG富集分析发现,不同碱处理时间下的KEGG途径(pathways)具有显著差异。对7个碱性胁迫反应相关基因进行RT-qPCR验证,所得基因的表达模式与RNA-Seq数据结果一致,证实了RNA-Seq结果的可靠性。本研究首次对碱胁迫下的榆叶梅叶片进行了转录组分析,获得的序列数据极大地丰富了榆叶梅可利用的基因资源,深入了解了榆叶梅碱胁迫的分子响应机制,同时为研究以榆叶梅为砧木的中国李(Prunus salicina)在碱胁迫下的适应机制奠定了分子研究基础。     

温度胁迫对幼体大海马基因转录表达的影响

为探究大海马幼体在高温和低温胁迫条件下基因表达水平的变化规律,采用Illumina平台的PE150测序功能对大海马幼体肝脏进行了转录组测序。对照组、高温胁迫组和低温胁迫组测序数据组装后获得22 513个单基因簇(Unigene),N50为2 223 bp,平均长度为1 122.71 bp。与对照组相比,高温胁迫组共获得14 009个差异表达基因,其中5 543个差异表达基因上调,8 466个差异表达基因下调;低温胁迫组共获得20 030个差异基因,14 016个差异表达基因上调,6 014个差异表达基因下调。差异表达基因经KEGG数据库富集发现,高/低温胁迫均导致大海马体内抗氧化途径相关基因表达(HSP70、HSP90、SOD等)发生显著改变;另外,高温胁迫还造成大海马免疫系统相关基因(PIK3R、Akt、IL-10、TLR等)以及细胞凋亡相关基因(CYCS、CASP9、CASP3等)表达显著改变;低温胁迫造成DNA损伤修复(MSH、DDB2、XRCC2、RAD52、Ogg1、PMS2等)、脂肪酸代谢(StARD7、ApoA4、CYP51、Fadsd6等)和细胞凋亡(P21、P53、BAX等)相关基因显著上调。选择HSP70、Fadsd 6与IL-10等13个差异基因验证,RT-qPCR结果与RNA-Seq测序结果基本一致。本研究结果为深入探究大海马温度应激的调控机制奠定了一定的理论基础,而且有助于预防极端温度对大海马造成损伤。     

油料作物油脂合成调控研究进展

植物油脂不仅是人类可食用油的主要来源,也是人类生产生活中重要的可再生原料。本文概述了植物油脂的生物合成途径,从母体效应、QTL、GWAS等多个方面总结了油料作物油脂合成的遗传学研究进展,同时探讨了已知的油脂合成调控相关基因的功能。本文综述了该领域的研究现状,为深入了解油料作物油脂合成调控网络提供了参考,也为油料作物的分子改良和遗传育种提供了理论基础。     

A novel glycoside hydrolase 74 xyloglucanase CvGH74A is a virulence factor in Coniella vitis

Grape white rot is a destructive fungal disease occurring worldwide. Recently, Coniella vitis was identified as the predominant pathogen causing this disease in China. As the periderms of grape shoots are severely degraded by C. vitis, it was speculated that cell wall-degrading enzymes (CWDEs) might play a key role in the pathogenesis of this disease. Therefore, this study aimed to examine the hydrolytic activity of the CWDEs of C. vitis. The results showed that xylanase (Xy) and xyloglucanase (XEG) had high levels of hydrolytic activity both in vitro and in vivo. Furthermore, a high-virulence fungal strain exhibited higher levels of Xy and XEG activities compared with a low-virulence strain. The genome of the fungus was found to harbor two XEG-coding genes CvGH74A and CvGH74B, which belonged to the glycoside hydrolase (GH)74 family. The expression level of CvGH74A was found to be high during pathogen infection. CvGH74A gene deletion mutants were generated using the split-marker method. The deletion of CvGH74A decreased both the hydrolytic activities of XEG and Xy and also the ability of the fungus to infect the grape leaves. No differences in the hyphal growth, morphology of colonies, or conidiation were found between the ΔCvGH74A mutant strains and the wild-type strain. Together, these results suggested that CvGH74A acted as an important virulence factor, and its enzymatic activity might regulate the virulence of the pathogen. This study was novel in reporting that GH74 XEG acted as a virulence factor in C. vitis.     

不同培养基继代培养蜡蚧轮枝菌对产孢量和烟粉虱毒力的影响

为了探究不同培养基继代培养的蜡蚧轮枝菌对烟粉虱的作用效果,通过继代培养方法测定JMC-01、JMC-02菌株的产孢量、对烟粉虱的毒力及其几丁质酶活性。结果表明,JMC-01、JMC-02菌株在添加蝉蜕和烟粉虱的PDA培养基中连续培养5代的产孢量、对烟粉虱的毒力、几丁质酶活性均高于无添加的PDA培养基,且JMC-01菌株的各项参数均高于JMC-02菌株。JMC-01菌株在添加蝉蜕的PDA培养基中T_1和T_3代的产孢量最大,均为1.279×10~7个/mL,T_3代对烟粉虱的毒力最大,为88.33%,T_5代的几丁质酶活最高,为0.268 3 U/mL;在添加烟粉虱的PDA培养基中,JMC-01菌株的最大产孢量、对烟粉虱的最高毒力和最高几丁质酶活分别出现在S_2、S_4和S_5代,分别为1.270×10~7个/mL、88.33%和0.325 4 U/mL。回归分析结果显示,JMC-01、JMC-02菌株几丁质酶活性与烟粉虱校正死亡率呈显著的正相关,且拟合优度较好。本试验表明JMC-01、JMC-02菌株均对烟粉虱具有较强的毒力,添加蝉蜕和烟粉虱的培养基可以维持菌株生长特性、增加产孢量、延缓菌株退化、增强菌株的几丁质酶活性,为烟粉虱的生物防治提供了新的菌种资源。     

Sodium alginate supplementation modulates gut microbiota,health parameters,growth performance and growth‐related gene expression in Malaysian Mahseer Tor tambroides

This study investigated the effects of sodium alginate supplementation on gut microbiota composition, health parameters, growth performances and growth‐related gene expression of Malaysian mahseer. Five test diets were formulated by supplementing 0%, 0.1%, 0.2%, 0.4% and 0.8% sodium alginate. Triplicate groups of juvenile Tor tambroides (2.19 ± 0.05 g) were stocked in 15 aquaria (20 individuals per aquarium) and fed at 3.0% body weight per day for 60 days. PCoA and UPGMA analysis showed that gut bacterial community were more convergence in higher sodium alginate‐supplemented diets. The percentage of Porphyromonadaceae, Bacteroides, Plesiomonas and Shewanella were substantially higher and Aeromonas, Entomoplasmatales and Prevotellaceae were drastically lower in higher sodium alginate (0.2%–0.8%) diets. Sodium alginate supplementation (≥0.2%) significantly improved the haematocrit value and respiratory burst activity of T. tambroides. Growth performances and feed utilization were significantly higher in 0.2%–0.4% sodium alginate‐supplemented diets. The increased growth rate of T. tambroides was governed by both hyperplastic and hypertrophic muscle growth. Real‐time PCR data demonstrated that most of the growth‐related genes were significantly upregulated in 0.2%–0.4% sodium alginate‐supplemented diet. Finally, it can be concluded that sodium alginate should be supplemented at 2 g/kg in practical fish feed formulation.     

Effects of five compound attractants in high plant‐based diets on feed intake and growth performance of juvenile turbot (Scophthalmus maximus L.)

An experiment was conducted to evaluate the effects of five attractants on feed intake, growth performance and appetite‐regulating genes expression for juvenile turbot (Scophthalmus maximus L.). Fish were fed with seven diets: fish‐meal‐based diet (FM), fermented soybean meal replacing 45% fish meal (FSBM) and other diets with 1% compound attractant A (CA‐A) consisting of amino acids, compound attractant B (CA‐B) consisting of quaternary ammonium bases, compound attractant C (CA‐C) consisting of nucleotides, compound attractant G (CA‐G) consisting of the combination of CA‐A, CA‐B and CA‐C and blue mussel protein hydrolysate (BMH) on the basis of FSBM diet respectively. Daily feed intake in CA‐A group was significantly higher, but that in CA‐B group was significantly lower than FSBM group. Similarly, specific growth rate was significantly enhanced in CA‐A and CA‐G groups and were significantly decreased in CA‐B group. Protein efficiency ratio and protein retention in CA‐B group were significantly lower than FSBM group. Furthermore, BMH supplement increased the mRNA expression of Ghrelin in the intestine of fish. These results showed that CA‐A and CA‐G supplement could enhance growth in plant‐based diets, but CA‐B supplement had the opposite effect, which might be associated with lower feed intake and lower feed utilization.     

二丁基羟基甲苯对雨生红球藻虾青素和油脂积累的影响

以雨生红球藻(Haematococcus pluvialis)为对象，研究高光照、缺氮条件下，外源添加不同浓度的二丁基羟基甲苯(Butylated hydroxytoluene, BHT)对雨生红球藻生长、虾青素积累、油脂合成、脂肪酸组成、碳水化合物、蛋白含量以及虾青素和脂肪酸合成相关酶基因的影响。结果显示，添加不同浓度的BHT后，2 mg/L BHT添加组虾青素积累量为最高，显著高于其他实验组和对照组(P<0.05)，达到31.66 mg/g，是对照组的1.87倍。油脂含量达45.56%，高于同期对照组(39.06%)，脂肪酸组成变化不显著。在此条件下，虾青素合成关键酶基因dxs和bkt表达水平分别是对照组的5.19倍和2.04倍；脂肪酸合成关键酶基因kas和acp表达水平较对照组显著提高(P<0.05)，分别是对照组的4.56倍和3.02倍。与对照组相比，2 mg/L BHT添加组的碳水化合物和蛋白含量均呈下降趋势。研究表明，在胁迫条件下，外源添加适量浓度的BHT能有效促进雨生红球藻中虾青素的积累，同时提高了藻细胞内的油脂含量。