石榴转录组密码子使用偏向性

对石榴(Punica granatum)转录组数据进行从头组装,预测完整CDS序列的密码子使用偏向性;将石榴转录谱与其他8种蔷薇分支物种基因组编码序列进行同义密码子相对使用度(RSCU)比较,并对石榴WRKY和MYB家族基因进行亚家族进化分析和RSCU比较。结果表明:(1)石榴转录组主要受自然选择压影响。密码子偏向性极强的基因多为细胞相关功能基因;没有密码子使用偏向性的基因多与维持生命活动本质相关。(2)基因差异高表达影响RSCU,CpG二核苷酸富集影响密码子适应性。(3)物种间亲缘关系越近,RSCU值越相似。对蔷薇分支物种密码子进行RSCU比较,石榴与巨桉(Eucalyptus grandis)偏向性基本一致。(4)亚家族功能分化影响密码子偏向性。石榴WRKY亚家族1、4和13在翻译精氨酸、亮氨酸时分别偏向使用AGG、CUC,这与其不断进化的抗逆功能相关。石榴MYB亚家族8在翻译苏氨酸时偏向选择ACA,这与其扩张中的木质化功能相关。     

草地贪夜蛾Sf9细胞对饥饿诱导自噬的响应

为研究草地贪夜蛾对饥饿诱导自噬的响应。从草地贪夜蛾EST数据库中筛选到一段核苷酸序列,设计特异性引物,RT-PCR技术从草地贪夜蛾Sf9细胞中扩增该核苷酸序列,经过生物信息学分析该序列与昆虫自噬相关基因ATG8同源性最高,命名为SfATG8基因,并构建了pIEX-4-mCherry-EGFP-SfATG8载体转染Sf9细胞。Sf9细胞经饥饿(PBS)诱导4 h后,Lyso-Tracker染色、Western Blotting、共聚焦显微镜检测细胞对自噬的响应程度。结果表明,经饥饿(PBS)诱导4 h的Sf9细胞中出现Sf Atg8-PE的表达,共聚焦显微镜检测到Sf9细胞膜上的自噬小体。以上证据证实饥饿(PBS)能诱导Sf9细胞发生自噬,为深入研究Sf9细胞对自噬的响应奠定了基础。     

日光温室蔬菜栽培废弃基质露地还田对土壤理化性质及辣椒生理特性和产量的影响

为了研究基质还田对大田土壤和辣椒生长生产的影响,以日光温室连续栽培蔬菜6年的废弃基质还施露地种植辣椒,测定了不同施用量(0,45,90,135 m~3/hm~2)的土壤理化性质及辣椒生理特性和产量。结果表明:基质还田能够减少土壤pH变化,减小土壤容重、略微增加土壤孔隙度、保持土壤气水比,改善了土壤的理化性状,促进辣椒植株的生长且利于其干物质积累,其中株高、茎粗及干物质积累量均以T2最好。提升了辣椒叶片光合荧光的表现,T2的净光合速率、蒸腾速率、气孔导度、Fv/Fm、ΦPSⅡ均为最高,T1的q P最高,NPQ为T3最低。提高了辣椒果实的品质及产量,T2产量达67.04 t/hm~2,可溶性糖含量以T2最高,为2.21%,Vc含量T3最高,为1.97 mg/g,T1的可溶性蛋白含量最高、硝酸盐含量最低,分别为2.07,204.93 mg/kg。而与此同时,较大的还田施用量(135 m~3/hm~2)在辣椒生长的中前期造成了一定的毒害现象,在生育后期此现象消失,最终各项指标全面优于未还田处理。废弃基质露地还田这样的重复利用方式切实有效,(在种植同科作物的情况下)还田施用量宜选择90 m~3/hm~2。     

抗白粉病胡麻种质资源田间鉴定与筛选

白粉病目前已成为影响胡麻产量和质量最常见的病害之一, 种植抗病品种是防控病害最经济环保的有效措施, 然而抗病亲本材料的缺乏已成为制约抗病品种选育的关键因素。为筛选出抗白粉病胡麻材料, 本研究在田间自然感病的条件下, 采用病情指数法对300份国内外胡麻种质资源材料进行了抗白粉病鉴定和评价。结果表明, 所有供试材料均程度不同地感染胡麻白粉病, 无免疫材料, 仅有5份材料为中抗; 其余295份均为感病材料, 其中8份材料中感, 52份材料感病, 235份材料高度感病。本研究可为抗白粉病胡麻品种的培育及相关抗病基因的发掘提供参考。     

一株溶磷菌的分离鉴定及溶磷促生作用

为了提高土壤磷素利用率,为微生物磷肥开发提供高效菌种资源,采用平板分离法从荒漠绿洲过渡带白刺根际土壤分离筛选出一株溶磷能力强的真菌P-1407,对其溶磷特性、产酸特性和耐盐碱特性进行研究。结果表明,液体条件下培养10 d,菌株 P-1407对 Ca₃( PO₄)₂表现出高效的溶解能力,最大溶磷量为557 μg·mL^-1;其溶磷量和pH值变化呈显著负相关,溶磷过程中分泌的有机酸种类主要为葡萄糖酸、草酸及甲酸;在高浓度 NaCl 和较高的pH值条件下仍保持较高的溶磷能力;结合培养性状、显微形态特征和ITS rDNA序列分析,鉴定菌株P-1407为草酸青霉（Penicillium oxalicum）;盆栽试验表明,P-1407菌株对番茄的促生作用较对照显著提高,番茄种子发芽率提高了2.53%,番茄植株的株高、根长、地上部鲜重、地上部干重、地下部鲜重和地下部干重分别增加了109.27%、78.41%、82.95%、121.69%、94.74%和64.71%。因此,P-1407是一株展现良好应用前景的高效溶磷菌种,有望为土壤的改良和生物菌肥的开发提供种质资源。     

吡咯伯克霍尔德氏菌JK-SH007产嗜铁素的相关基因克隆及条件优化

为了明确吡咯伯克霍尔德氏菌Burkholderia Pyrrocinia JK-SH007是否产嗜铁素,揭示其生防机制,本文通过CAS检测该菌株产嗜铁素的能力,并对其嗜铁素合成相关基因cepR进行克隆及序列分析,同时采用单因素试验及响应面法对菌株JK-SH007产嗜铁素条件进行优化。结果表明,菌株JK-SH007具有明显的产嗜铁素能力,其嗜铁素合成相关基因cepR大小为720 bp,与B.pyrrocinia（EU034001）的亲缘关系最近,同源性为99%,两者序列相似性为97%,13个位点出现SNP;另外菌株JK-SH007 cepR基因编码的氨基酸序列有3个氨基酸发生了替换,两者一致性为97%。该菌产嗜铁素最优培养时间是15 h、pH 8、转速200 r/min、最佳碳源和氮源分别是甘油和氯化铵;PB试验筛选出影响该菌株产嗜铁素的关键性因素分别是pH、加碳量、加氮量;CCD试验结果显示,pH和加氮量的交互作用较明显;最终采用Design-Expert软件分析出菌株JK-SH007产嗜铁素最佳方案为碳源加入量15.00 g/L、氮源加入量10.50 g/L、温度30℃、pH 7.36,优化后菌株产嗜铁素能力由18.59提升到37.86,响应面优化产嗜铁素条件的效果是显著的,嗜铁素产量得到明显提高。     

群体感应信号物质对吡咯伯克霍尔德氏菌JK-SH007在杨树内定殖的影响

为了明确杨树溃疡病生防菌吡咯伯克霍尔德氏菌JK-SH007的群体感应系统是否与其内生定殖相关,本文通过群体感应指示菌紫色杆菌CV026和液相色谱串联四级杆飞行时间质谱技术(HPLC-Q-TOF-MS)确定其群体感应信号物质种类,并利用结晶紫染色、菌体回收、GFP标记等技术,研究其群体感应信号物质对该菌株生物膜及在杨树苗内的定殖能力的影响。结果表明,菌株JK-SH007产生的群体感应信号物质为含8个碳原子酰基侧链的辛酰基-L-高丝氨酸内酯(C8-HSL)。群体感应信号物质C8-HSL的添加对该菌株生物膜及在杨树苗内的定殖能力有影响,并且呈现低浓度促进,较高浓度抑制的规律。C8-HSL的添加与菌株JK-SH007生物膜的OD值和菌体量具有显著相关性,相关系数分别为0.923和0.756。当添加1.0%C8-HSL终浓度达到5×10^-8 g/L时,菌株JK-SH007生物膜的形成量达到峰值。荧光显微镜镜检发现,当添加100μL C8-HSL时,杨树组培苗根和茎中GFP荧光标记的菌株JK-SH007的定殖数量明显较CK多。菌株JK-SH007的群体感应与其在杨树内的内生定殖能力密切相关,群体感应信号物质C8-HSL具有增强菌株JK-SH007内生定殖的能力,同时也表明该物质有利于该菌株生防效果的发挥。     

低温驯化对南方小花蝽冷藏的影响

为探讨低温驯化对南方小花蝽雌成虫冷藏的影响,在室内15℃下饲养7 d作为低温驯化处理,研究了驯化后与未驯化的南方小花蝽雌成虫分别在4、6和10℃下冷藏其寿命、存活率和致死中时间（LT₅₀）的异同。结果表明,无论是否进行低温驯化,南方小花蝽雌成虫的寿命都显著大于对照（25℃下饲养）。在10℃冷藏条件下,是否经过低温驯化对南方小花蝽雌成虫寿命没有明显影响,但在4℃和6℃条件下,低温驯化能明显延长雌成虫的寿命,分别延长了80.7%和83.7%。南方小花蝽雌成虫存活率均随冷藏时间的延长而下降,在同一低温冷藏温度下经低温驯化处理的南方小花蝽存活率比未驯化的高。经低温驯化的南方小花蝽雌成虫在4、6和10℃冷藏时,其LT₅₀分别比未经驯化的延长了30.0%、57.8%和40.4%。以上结果表明南方小花蝽的寿命具有可塑性,低温驯化有利于南方小花蝽的冷藏,6℃可作为南方小花蝽长期储藏的温度。     

Ellagic acid attenuates hypoxic-ischemic brain injury by alleviating autophagy

AIM:To investigate whether ellagic acid (EA) attenuates hypoxic-ischemic encephalopathy (HIE) by down-regulating autophagy. METHODS:In vivo, Sprague-Dawley rats (n=17) were randomly divided into 3 groups:5 rats for sham group, 6 rats for HIE group and 6 rats for HIE+EA pretreatment group. The rats in HIE+EA pretreatment group were treated with EA (10 mg/kg, 10 mL/kg, suspended in corn oil, ig). After 24 h of operation, the rats from each group were sacrificed and their brains were collected. TTC staining and HE staining were used to define the infarct areas and brain structure. The autophagy-related proteins beclin-1, P62, LC3-Ⅱ/-I and Atg5 in the cortex in each group were compared by Western blot. In vitro, PC12 cells were divided into 3 groups:control group, CoCl₂ group and CoCl₂+EA pretreatment group. CoCl₂ at 800 μmol/L was added to the PC12 cells to induce an anoxic environment. The PC12 cells were pretreated with EA at 8 μmol/L and the cell viability was measured by CCK-8 assay. The production of reactive oxidative species (ROS) in the cells was detected by flow cytometry with DCFH-DA staining. MDC staining and TMRE staining were applied to reflect the extent of autophagy and the state of apoptosis, respectively. The autophagy-related proteins in PC12 cells were also investigated. RESULTS:In HIE group, 7-day-old rats were given the operations and the their large infarct areas in the hemisphere were observed by TTC staining. HE staining displayed the injured hemispheres which contained few neurons, and exhibited edema status and serious structural damage. EA pretreatment decreased the infarct area and alleviated the damage to hemisphere with more visible neurons, compared with HIE group. Compared with sham group, the levels of autophagy-related proteins Atg5, beclin-1 and LC3-Ⅱ/-I in the cortex were increased (P<0.01), and P62 protein expression was decreased (P<0.01) in HIE group. Compared with HIE group, the protein expression of Atg5, beclin-1 and LC3-Ⅱ/-I was decreased (P<0.01) and P62 protein expression was increased in HIE+EA pretreatment group (P<0.01). In vitro, compared with CoCl₂ group, the PC12 cells in CoCl₂+EA pretreatment group showed a lower ROS level. Moreover, the cells in CoCl₂+EA pretreatment group exhibited higher mitochondrial membrane potential than that in CoCl₂ group. MDC staining in CoCl₂ group showed high value of fluorescence and increased number of autophagosomes. EA pretreatment reduced the number of autophagosomes and the extent of autophagy to protect PC12 cells. Furthermore, the protein levels of Atg5, beclin-1 and LC3-Ⅱ/-I in CoCl₂ group were higher (P<0.01), and the protein expression of P62 was lower (P<0.01) than those in control group. In CoCl₂+EA pretreatment group, the protein levels of Atg5, beclin-1 and LC3-Ⅱ/-I were decreased (P<0.01) and the protein expression of P62 was increased as compared with CoCl₂ group (P<0.01). CONCLUSION:EA pretreatment attenuates autophagy to protect the neurons against HIE injury.