几种经济红藻中茉莉酸合成途径的关键物质

为研究几种经济红藻中茉莉酸合成途径的关键物质,实验采用液相色谱—质谱联用技术(LC-MS)对茉莉酸生物合成途径相关物质建立检测方法,并对龙须菜、坛紫菜受到机械损伤时各物质的变化情况进行检测。以100%甲醇提取茉莉酸(JA)、茉莉酸甲酯(MeJA)、12-氧-植物二烯酸(12-OPDA)和13-氢过氧化亚麻酸(13-HpOTE),利用XBridge TM C18色谱柱,以甲醇/水为流动相分离4种物质。优化条件下4种物质得到良好分离,加标回收率为81.23%～90.25%,检测限为0.04～0.56 ng/mL,灵敏度高。坛紫菜、龙须菜和真江蓠3种红藻中,坛紫菜中未检测到JA和13-HpOTE,4种物质均在另外2种藻中检测到。龙须菜受机械损伤胁迫后,4种物质在短时间内得到积累,其中13-HpOTE响应迅速。研究表明,红藻中可能存在类似于植物的茉莉酸合成途径,并参与对损伤的胁迫响应。     

连栽木麻黄根际微生物群落结构和功能特征

以不同栽培代数的木麻黄(第1代FCP、第2代SCP、第3代TCP)根际土壤为试验材料,运用BIOLOG微平板和磷脂脂肪酸(PLFA)技术分析根际土壤微生物群落结构和功能多样性对多代连栽响应。结果表明,不同代数的土壤微生物对各类碳源的利用程度存在显著差异,连栽后木麻黄根际土壤微生物对碳源利用率显著下降。在6类碳源中,除胺类外,其他5种碳源均呈现FCP>SCP>TCP。PLFA分析共检测到11种PLFA生物标记,FCP土壤微生物PLFA生物标记总量明显高于SCP和TCP,3个代数土壤中含量最高的PLFA生物标记是i16:0、a15:0和18:1ω9c。土壤中特征微生物含量差异明显,细菌分布量最大,其次是真菌和放线菌。随栽植代数增加,细菌含量减少,真菌含量增加。土壤微生物群落多样性指数均呈现FCP>TCP>SCP,与土壤理化性质变化密切相关。可见木麻黄连栽显著影响其根际土壤微生物群落结构与功能,因此根际土壤微生态失衡可能是导致木麻黄连栽障碍的重要因素。     

胡麻脂肪酸去饱和酶基因(fads)差异表达分析

为了了解fad基因在胡麻蒴果发育过程中对不饱和脂肪酸的调控,对高、中、低三个不同亚麻酸(C18:3)含量的胡麻品种(’CDC Gold’,‘内亚7号’,’Linola’)进行了不同时期的品质测定,以及脂肪酸去饱和酶2a基因(fad2a)、脂肪酸去饱和酶2b基因(fad2b)、脂肪酸去饱和酶2c基因(fad2c)、脂肪酸去饱和酶3a基因(fad3a)、脂肪酸去饱和酶3b基因(fad3b)的qRT-PCR定量分析。结果表明,随着蒴果成熟,可溶性糖含量呈降低趋势,粗脂肪与粗蛋白不断积累,且差异显著(p<0.05)。fad2a基因、fad3a基因以及fad3b基因在各个时期中的表达符合正态分布。以0 d的蒴果为对照,在胡麻种子形成过程中,‘内亚7号’的三个基因在5 d和15 d的表达量迅速增加,到30 d时急剧减少,15 d的fad2a基因表达量为5.23倍,fad3a基因表达量是fad2a基因表达量的14.52倍,fad3b基因的表达量是fad2a基因表达量的16.14倍,表明这三个基因参与不饱和脂肪酸积累过程。在低亚麻酸含量品种‘Linola’30 d中,fad3a基因是fad2a基因表达量的52.71倍,fad3b呈下调趋势;在高亚麻酸含量品种‘CDCGold’30d中,fad3a基因与fad2a基因的表达量均呈下调趋势,fad3b的表达量为3.92倍;在中等亚麻酸含量品种‘内亚7号’中,fad2a基因表达量降低了0.31倍,fad3a基因是fad3b基因表达量的1.87倍。fad2b基因、fad2c基因熔解曲线不稳定,峰值低,可以在后续试验中继续探索。     

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.     

Crude protein in low‐fish meal diets for juvenile Litopenaeus vannamei can be reduced through a well‐balanced supplementation of essential amino acids

The use of supplemental essential amino acids (EAAs) has been shown to provide an opportunity to minimize excess levels of crude protein (CP) in animal feeds. The present study investigated the effect of reducing the amount of CP in low‐fish meal diets (5%) for juvenile Litopenaeus vannamei. Four sets of diets were prepared containing (% on a fed basis, mean ± SD) 31.24 ± 0.71, 33.70 ± 0.41, 36.90 ± 0.44, and 39.63 ± 0.14% CP, with each protein level containing a total dietary methionine (Met) (Met + Cys) level of 0.56 ± 0.02 (1.07 ± 0.23), 0.71 ± 0.01 (1.22 ± 0.20), 0.88 ± 0.02 (1.38 ± 0.22), or 1.04 ± 0.02% (1.55 ± 0.18%). Shrimp of 1.00 ± 0.08 g were stocked in 84 outdoor tanks of 1 m³ at a rate of 100 shrimp/m² and raised for 75 days. Final survival ranged from 83 to 97% and was unaffected by Met content. Both survival and yield were significantly depressed when shrimp were fed the 31% CP diet. Shrimp grew at a weekly rate of between 0.79 and 0.97 g, achieving a final body weight (BW) in excess of 10.8 g. There was a significant interaction between CP and Met over BW. Shrimp fed 0.56% Met achieved the lowest BW at harvest. Increasing CP beyond 34% did not enhance BW. With a dietary Met content of 0.71%, the highest BW was achieved with 34% CP compared to other levels. There was a significant improvement in Feed Conversion Ratio (FCR) when CP was raised from 31 to 34%. Similarly, dietary Met levels above 0.71% resulted in a significantly better FCR compared to 0.56%. Our study has shown that, if dietary Met (Met + Cys) meets a minimum of 0.71% (1.22%), levels of CP could be reduced from 40 to 34% without adverse effects on shrimp performance.     

氨基酸配合硼喷施对油麦菜硼营养及生长、品质的影响

为探究有机营养类物质氨基酸与硼酸配合喷施对油麦菜生长的应用效果,本研究采用盆栽试验,设置单独喷施1%硼酸处理,以及谷氨酸、丙氨酸和天冬氨酸3种氨基酸各设3个喷施浓度(5、10、20 mmol·L^-1) 与1%硼酸配合喷施处理,并以喷施清水为对照(CK),共11个处理,收获后测定植株的生长指标和硼含量。结果表明,单独喷施硼酸对油麦菜生物量无明显影响,低浓度氨基酸与硼配合喷施均能显著提高油麦菜生物量,其中丙氨酸-硼喷施处理油麦菜生物量较CK平均增加了9.4%。与CK相比,3种氨基酸与硼配合喷施均能明显提高油麦菜叶片总蛋白含量,同时降低硝酸盐积累,从而改善品质。与单独喷施硼酸相比,适宜浓度氨基酸与硼配合喷施均可显著提高油麦菜氮、钾积累量,另外谷氨酸-硼和天冬氨酸-硼喷施对油麦菜磷积累量也有明显促进作用。与单独喷施硼酸相比,氨基酸与硼配合喷施均可显著提高油麦菜地上部硼含量,其中丙氨酸-硼喷施处理油麦菜地上部硼含量平均增幅为41.0%;此外,丙氨酸-硼喷施对油麦菜地下部硼含量也有明显提升作用,平均增幅为15.6%;喷施丙氨酸浓度与油麦菜地上部和地下部硼含量均呈显著正相关关系。综合比较不同氨基酸与硼酸配合喷施对油麦菜的影响,可考虑将丙氨酸作为增效剂与硼肥配合施用,提高硼肥的应用效应。本研究为开发新型硼肥提供了理论依据。     

草酸处理对桂七芒果冷害及细胞壁代谢的影响

为探讨草酸处理对低温冷藏下芒果冷害及细胞壁代谢的影响,本试验以桂七芒果果实为试材,采用5 mmol·L^-1草酸溶液浸泡处理,以清水浸泡处理为对照,并于4℃贮藏,分析芒果的冷害指数、丙二醛(MDA)含量、相对电导率、硬度、细胞壁物质含量、细胞壁代谢酶的变化。结果表明,与对照组相比,草酸处理显著降低了低温贮藏14 d后桂七芒果果实冷害指数、MDA含量、相对电导率、原果胶和纤维素含量,显著降低了低温贮藏28 d后果实的硬度;显著提高了低温贮藏14 d后果实水溶性果胶含量及多聚半乳糖醛酸酶(PG)、果胶甲酯酶(PME)、纤维素酶(Cx)活性,显著提高了贮藏21 d后果实β-半乳糖苷酶(β-Gal)活性。综上所述,草酸处理能减轻桂七芒果冷害,维持采后果实细胞壁降解酶较高活性和水溶性果胶含量。本研究为揭示草酸减轻芒果果实冷害机制提供了依据,可为草酸应用于其他冷敏型果实的贮藏保鲜提供理论参考。