Napin启动子特异性表达油菜BnGPAT9基因

甘油三磷酸酰基转移酶(GPAT)基因编码参与植物油脂生物合成的酶。为了提高油菜籽含油量,促进油菜增产增油,改善菜籽油脂肪酸组分,克隆甘蓝型油菜Napin启动子与三酰甘油合成相关基因BnGPAT9,构建pBI121-Napin-BnGPAT9植物过表达载体,利用农杆菌介导法转化拟南芥,采用气相色谱法分析种子中脂肪酸组分,通过索式抽提法测定种子中油脂含量,研究在种子中过表达BnGPAT9基因对种子油脂合成的影响。Napin启动子在线分析表明,克隆到的启动子具有大量TATA-box、CAAT-box以及ABRE、GCN4等特征元件,完全具备种子特异性表达功能;种子中脂肪酸组分分析显示,拟南芥种子中油酸(C18∶1)含量提高0.70～1.01百分点,亚油酸(C18∶2)和芥酸(C22∶1)含量降低;拟南芥种子含油量测定结果显示,种子中含油量显著增加1.91～2.56百分点。综上,甘蓝型油菜BnGPAT9基因对植物油脂合成具有重要作用,利用Napin启动子在拟南芥种子中过表达BnGPAT9基因可提高种子含油量并改善脂肪酸组成成分,为BnGPAT9基因应用于油菜油脂改良方面打下一定基础。     

食用菌创意景观设计

通过对食用菌创意景观设计进行研究,分析具体的设计原则和设计方法,在食用菌创意景观打造过程中融入艺术技巧和文化特色,营造独特的食用菌旅游观光资源,创造更多农业生产附加价值。     

快速灵敏的甘蔗线条花叶病毒免疫学检测体系的建立

 甘蔗花叶病广泛存在于我国甘蔗种植区,严重影响甘蔗产业的高质量发展。近年来甘蔗线条花叶病毒在蔗区肆虐,尽管针对其的血清学检测技术已经建立,但是快速、准确、高通量的检测方法亟待发掘。本研究制备了^SCSMVCP的抗血清,特异性高,与引起甘蔗花叶病的另两种病原 (高粱花叶病毒和甘蔗花叶病毒) 间没有血清学交叉反应。基于该多克隆抗体,建立了直接抗原包被的ELISA、斑点杂交、Western blot和基于多抗的免疫试纸条检测技术。开发的免疫试纸条检测技术能快速、准确、高通量应用于田间病毒鉴定。本文提供了基于血清学的快速、准确、高通量,且便捷的甘蔗线条花叶病毒检测技术,有助于我国蔗区甘蔗花叶病的监测与防控。     

刺萼龙葵种子中适宜内参基因的筛选

入侵杂草刺萼龙葵Solanum rostratum Dunal传播扩散的主要载体是种子,研究其种子休眠萌发基因的激素调控对于其防除具有重要意义,而选择合适的内参基因可以提高相关基因表达分析的准确性。本研究以赤霉素、脱落酸和水处理的刺萼龙葵种子为材料,利用geNorm、NormFinder、BestKeeper和RefFinder 4种软件对15个候选内参基因进行表达稳定性评价,并通过检测ABI5(abscisic acid-insensitive 5)的表达验证所筛选的内参基因的适用性。结果表明,对于赤霉素、脱落酸和水处理过的种子,最稳定的内参基因分别为eIF(eukaryotic initiation factor)、SAND(SAND protein family)和ACT(β-actin);对所有种子样本而言,PP2Acs(a catalytic subunit of protein phosphatase 2A)是最稳定的内参基因。研究结果将为刺萼龙葵种子休眠萌发的遗传调控研究提供重要参考。     

甘薯基因组NBS-LRR类抗病家族基因挖掘与分析

NBS-LRR类基因家族是植物抗病R基因(Resistance gene)数量最多的一类,具有NBS (Nucleotide-binding site)和LRR (Leucine-leucine-repeat)结构域。甘薯(Ipomoea batatas)栽培种基因组已完成测序,但尚未注释,本研究对甘薯基因组序列进行外显子预测,得到甘薯染色体组全基因组蛋白序列,在此基础上进一步对NBS-LRR家族基因鉴定和分析表明,甘薯基因组中含有379个NBS-LRR家族基因,占全基因组基因总数的0.212%,其中N型亚家族120个,NL型103个, CNL型133个, TNL型22个, PN型1个。所有染色体上均有NBS-LRR家族基因分布,但数量明显不同,其中有60.9%的NBS-LRR基因序列呈簇状分布。NBS-LRR基因序列有15个保守结构域,在N端较为保守。研究结果为甘薯进一步开展NBS-LRR家族基因的功能研究和抗性育种提供了参考。     

Rapid mapping of candidate genes for cold tolerance in Oryza rufipogon Griff. by QTL-seq of seedlings

Cold stress is a major problem in rice production. To rapidly identify genes for cold tolerance in Dongxiang wild rice(DWR, Oryza rufipogon Griff.), sequencing-based bulked segregant analysis of QTL-seq method was used to resequence the extremely resistant(R) and susceptible(S) bulks of a backcross inbred lines(BILs) population(derived from Oryza sativa×O. rufipogon) and their parents. Single nucleotide polymorphisms(SNP)-index graphs and corresponding Δ(SNPindex) graphs(at 99 and 95% confidence levels) for R-and S-bulks detected a total of 2 609 candidate SNPs, including 58 candidate cold-tolerance genes. Quantitative real-time PCR analysis revealed that 5 out of the 58 candidate genes had significant differences in expression between O. sativa and O. rufipogon. Structural variation and functional annotations of the 5 candidate genes were also analyzed, and allowed us to identify 2 insertion-deletion(InDel) markers(12-7 and 12-16) that were linked with candidate genes on chromosome 12 in DWR. These results are helpful for cloning and using cold tolerance genes from common wild rice in cultivated rice.     

土壤电处理技术防治黄瓜土壤根结线虫试验

以辽宁省大连市炮台镇黄瓜种植区为对象，研究了土壤电处理对土壤中造成土壤连作障碍的根结线虫杀灭情况。结果表明，延长通电时间可以有效提高电处理杀灭土壤中根结线虫的效果，延长通电时间至6 min，土壤中和根际的根结线虫数量已趋近于零，且经过电处理后的黄瓜每株数量和产量均有提升。      

温度对草地贪夜蛾生长发育及繁殖的影响

草地贪夜蛾是新近入侵我国的重大农业害虫,为了弄清其入侵生物学特性,为草地贪夜蛾的监测预警及综合治理提供理论依据,本研究在光照培养箱内设置6个温度(17、22、27、32、37、42℃)饲养草地贪夜蛾,测定其不同阶段的存活率、发育历期、体重及繁殖力等,探究温度对草地贪夜蛾生长发育及繁殖的影响。结果表明,草地贪夜蛾在42℃条件下仅能短暂存活,在17～32℃范围内可正常生长发育,其幼虫期、蛹期及世代历期均随着温度的升高而缩短,17℃条件下世代历期和幼虫期分别为(58.73±1.46)d和(21.79±1.03)d,37℃条件下则分别仅为(22.57±0.84)d和(11.12±0.31)d。成虫寿命在17～27℃范围内差异不大,之后随温度升高而缩短,37℃下成虫寿命仅为27℃下的1/4左右,且在该温度下成虫无法正常羽化,雌成虫不产卵。17～32℃下雌成虫产卵前期随温度的升高而缩短,产卵期则随温度升高而有所延长,平均单雌产卵量在27℃下最高,为(565.50±143.40)粒。说明温度升高会导致草地贪夜蛾发育历期缩短,年发生世代数增加,幼虫取食量增大,对作物的为害加重。     

Effect of beclin-1 gene silencing on TuAKe-injured gastric cancer SGC-7901 cells

AIM: To observe the effect of beclin-1 silencing by the technique of RNA interference on the injury of human gastric cancer SGC-7901 cell by Sheliugu extract (the extract from tuber of Amorphophallus konjac, TuAKe). METHODS: To knock down the expression of beclin-1 gene, SGC-7901 cells were transfected with lentiviral vector carrying beclin-1-shRNA. The beclin-1 gene knock-down and non-knock-down SGC-7901 cells were treated with TuAKe. The cell viability was analyzed by CKK-8 assay. The percentages of apoptotic cells were detected by flow cytometry. The expression of beclin-1 and LC3 was detected by Western blot. RESULTS: The beclin-1 gene silencing decreased the protein expression of beclin-1 and increased the protein expression of LC3 in the SGC-7901 cells, leading to the decrease in cell viability and the increase in apoptotic rate (P<0.05). TuAKe increased the protein expression of beclin-1 and LC3 in the SGC-7901 cells, and decreased the protein expression of LC3 in the SGC-7901 cells with beclin-1 gene silencing, thus inhibiting the cell viability and increasing the apoptotic rate (P<0.05). CONCLUSION: Beclin-1 gene silencing inhibits the activation of beclin-1-related signaling pathway in gastric cancer SGC-7901 cells, and aggravates the injury of cell viability induced by TuAKe.     

我国蔬菜作物耐低温性研究进展

低温胁迫是蔬菜生产过程中的主要逆境因子，对蔬菜作物的生长发育、商品产量形成影响极大。本文综述了蔬菜作物耐低温性的遗传模型、生理调节和分子机制，展望了蔬菜作物耐低温性的研究前景。