爱媛28’香橙砧与枳砧在南平市引种表现与评价

2017年引进早熟杂柑品种‘爱媛28’香橙砧和枳砧苗分别在大棚设施和露天进行试种，栽培4年调查比较两个砧木组合的生长表现、丰产性与果实品质，结果表明：‘爱媛28’香橙砧树表现生长势强，冠幅、树高、茎粗、结果枝、秋梢等生长量指标显著大于枳砧树，3a树初产1436 kg/667㎡，4a树2523kg/667㎡，显著高于枳砧4a树初产1809kg/667㎡；大棚种植香橙砧4a树果实10月下旬TSS11-12%，12月中旬TSS可达15%，与枳砧相当，可滴定酸比枳砧略高，两者固酸比均达18以上，口感风味极佳，两者单果重、果形指数、果皮厚度、可食率无显著性差异；未密封大棚4a树体经受2020年寒冬极端低温考验无明显冻害，抗旱抗寒能力中等，易感黑点病和流胶病，大棚种植能很好预防黑点病。综合评价爱媛28枳砧和香橙砧均适合在南平市种植，香橙砧比枳砧早期丰产性稳产性好，收益更高，香橙砧‘爱媛28’设施种植较枳砧更具优势。     

不同水肥条件下夏玉米/冬小麦农田生态系统碳平衡研究

农田生态系统碳平衡取决于农作物固定碳量和土壤异养呼吸排放碳量。为揭示水肥用量对农田生态系统碳平衡的综合影响，设置3个灌水水平：高水、中水和低水（W1、W0.85、W0.7夏玉米季分别为90、76.5、63mm，冬小麦季分别为140、119、98mm），4个施氮水平：高氮、中氮、低氮和不施氮（N1、N0.85、N0.7、N0夏玉米季分别为300、255、210、0kg/hm2，冬小麦季分别为210、178.5、147、0kg/hm2），4个施磷水平：高磷、中磷、低磷和不施磷（P1、P0.85、P0.7和P0夏玉米季分别为90、76.5、63、0kg/hm2，冬小麦季分别为150、127.5、105、0kg/hm2）进行了田间试验。结果表明：不同水肥处理下夏玉米/冬小麦农田生态系统表现为碳汇，夏玉米季净生态系统生产力固碳量（CNEP）为6805~7233kg/hm2，冬小麦季CNEP为5842~6434kg/hm2，夏玉米CNEP高于冬小麦。在高、中、低肥水平下，增加灌水量，夏玉米/冬小麦周年净初级生产力固碳量（CNPP）提高2.48%~5.96%，土壤微生物异养呼吸碳释放量（CRm）增加2.15%~15.20%，净生态系统生产力固碳量（CNEP）增加1.16%~6.47%。在高、中、低供水水平下，增加施肥量，夏玉米/冬小麦周年CNPP增加2.95%~3.43%，土壤CRm增加5.23%~18.67%，CNEP增加0.93%~2.79%，CNEP增加比例与供水水平呈负相关。在低水条件下，氮磷肥配施处理夏玉米/冬小麦农田周年CNEP较单施氮、磷肥分别增加4.86%、7.34%，且氮磷肥交互作用显著（P<0.05），水肥供应水平相差15%时对冬小麦农田CNEP有显著的正交互作用。氮磷肥配施、水肥协调供应均有助于促进夏玉米/冬小麦农田生态系统的净碳输入，在节水节肥原则下，夏玉米和冬小麦分别在W0.85N0.85P0.85和W0.7N0.85P0.85水肥供应条件下有利于增加农田CNEP。     

CnAβ基因对大鼠RBL-2H3细胞脱颗粒的影响

过敏是人和养殖动物常见的疾病，而且发生的频率呈现逐渐增加的趋势。为了深入了解过敏反应的机制，本研究利用CRISPR/Cas9基因编辑技术，构建钙调磷酸酶A beta （calcineurin A beta，CnAβ）基因敲除的大鼠RBL-2H3细胞株，并探讨CnAβ基因对RBL-2H3细胞生长及脱颗粒的影响。选取大鼠CnAβ基因第一外显子为敲除靶点，设计并合成3个单导向RNA （single guide RNA，sgRNA），构建pX459-CnAβ-sgRNA质粒，并用脂质体3000将构建好的质粒转染到RBL-2H3细胞内；利用嘌呤霉素对转染细胞进行筛选，通过DNA测序验证获得CnAβ基因敲除的RBL-2H3细胞株；并检测CnAβ基因缺失对细胞增殖和脱颗粒的影响。结果表明，成功构建了CnAβ单基因敲除的大鼠RBL-2H3细胞株；CnAβ基因缺失对RBL-2H3细胞增殖、细胞的颗粒形成以及颗粒含量无显著影响，但显著抑制由细胞表面受体介导的RBL-2H3细胞脱颗粒作用。该研究结果有助于深入了解动物过敏性疾病的发生机制，为动物过敏性疾病的预防提供了理论基础。     

Shikonin inhibits neuronal apoptosis induced by OGD through targeting PI3K/Akt signaling pathway

AIM: To explore the effect of shikonin on rat primary cortical neurons in oxygen-glucose deprivation (OGD)-induced injury model.METHODS: The neurons were pretreated with shikonin at different concentrations (0.02, 0.2, 2 and 20 μmol/L) followed by treatment with OGD. Lactate dehydrogenase (LDH) release assay and fluorescein diacetate/propidium iodide (FDA/PI) double staining were used to detect neuronal viability and apoptosis, and then the optimal concentration of shikonin was determined. LY294002 (PI3K/Akt signaling pathway inhibitor, 1 μmol/L) was added before the addition of shikonin, and the protein level of p-Akt (Ser473) in the neurons was determined by Wes-tern blot. LDH release assay and FDA/PI double staining were also used to detect neuronal viability and apoptosis.RESULTS: A certain concentration (0.2~20 μmol/L) of shikonin increased the viability of impaired neurons (P<0.05) and the protein level of p-Akt (Ser473) in the neurons (P<0.05). The effect of shikonin on neuronal p-Akt (Ser473) levels and the cell death were blocked by LY294002 (P<0.05).CONCLUSION: A certain concentration of shikonin reduces OGD-induced apoptosis of rat primary cortical neurons by activating PI3K/Akt signaling pathway.     

坛紫菜叶状体的抗生素联合除菌效果

为研究坛紫菜及其外生菌群对氯霉素、竹桃霉素和头孢噻肟3种抗生素的敏感性,通过将其与氨苄青霉素、卡那霉素和庆大霉素的组合,优化坛紫菜叶状体的除菌方法,并利用涂布平板、qRT-PCR及16S rRNA测序分析其除菌效果。结果显示,当坛紫菜先以氨苄青霉素(300 mg/L)、卡那霉素(100 mg/L)和庆大霉素(100 mg/L)混合处理18 h,再以氯霉素(50 mg/L)、头孢噻肟(200 mg/L)和竹桃霉素(50 mg/L)混合处理4 d,叶状体的健康率保持在96.3%以上,而对可培养细菌的抑制率达到99.9%。根据qRT-PCR和16S rRNA的结果可知,该种组合方法较前3种抗生素的组合,其菌总数下降41.5%,微生物的丰度和多样性指数明显下降。其中,假单胞菌、交替赤杆菌、交替单胞菌以及海杆菌等得到针对性的抑制。研究表明,多种抗生素的优化组合能够对坛紫菜叶片的菌群结构产生显著影响,使其受到抑制,但仍无法达到绝对除菌的效果。     

Agrobacterium-mediated transformation of the CrDAT gene and selection of transgenic periwinkle lines with a high vincristine accumulation

Catharanthus roseus contains vincristine and vinblastine, which are outstanding drugs for cancer. In the biosynthetic pathways of terpenoid indole alkaloids (TIAs) in C. roseus, deacetylvindoline 4-O-acetyltransferase (DAT) is a key enzyme that catalyses the last reaction of vindoline biosynthesis to form vinblastine and vincristine. In this study, the CrDAT transgene was transferred into the periwinkle by Agrobacterium-mediated transformation and generated transgenic periwinkle lines with an increase in vincristine accumulation. The C. roseus DAT gene was introduced into C. roseus plants and it was confirmed that CrDAT was successfully transferred into the genome of periwinkle plants and efficiently translated to synthesise recombinant DAT protein. Four transgenic periwinkle lines in T1 generation, T1-1, T1-3, T1-6, and T1-7, expressed recombinant DAT protein with the total protein content in the range of 2.86 μg.mg^?1 to 5.12 μg.mg^?1. Moreover, the vincristine contents of four transgenic lines increased by 1.63?2.48-fold compared to non-transgenic plants, ranging from 6.91 µg.g^?1 (fresh weight) to 10.53 µg.g^?1 (fresh weight). The T1-1 line had the highest vincristine content. Hence, the overexpression of the recombinant DAT protein can improve the vincristine accumulation of transgenic C. roseus plants.

Abbreviation: CrDAT - Catharanthus roseus Deacetylvindoline-4-O-Acetyl Transferase; D4H - Deacetoxyvindoline 4-hydroxylase; ELISA - Enzyme-Linked Immunosorbent Assay Monoterpene indole alkaloid; T0, T1 - Generations of transgenic plants; TIAs - Terpenoid indole alkaloids; WT- The wild-type tobacco plants (non transgenic plant); 35S - Cauliflower mosaic virus 35S promoter     

一个CRISPR/Cas9-VQR基因编辑系统的构建

CRISPR/Cas9系统是一种广泛应用于细菌、酵母、动物和植物中的基因组定点编辑技术,但该编辑系统的使用范围受PAM (proto-spacer-motif)位点NGG的限制。本研究通过突变Streptococcus pyogenes Cas9 (SpCas9)编码氨基酸(1135位的天冬氨酸D突变成缬氨酸V, 1335位的精氨酸R突变为谷胱氨酸Q, 1337位的苏氨酸T突变为精氨酸R,命名该突变子为Cas9-VQR)改造其识别PAM为NGA的位点以扩大其使用范围。并使用玉米Ubi启动子启动Cas9-VQR基因、优化SpCas9的密码子、加入保守的核定位信号序列、增加单子叶植物中保守的3′UTR序列和使用水稻U6启动子启动gRNA来修饰该编辑系统。结果表明Cas9-VQR系统能够识别PAM为NGA的位点,并进行有效的切割。体外酶切活性检测结果表明Cas9-VQR的切割效率为5%～70%。水稻转化检测结果表明Cas9-VQR的切割效率约为27.5%～70.5%,平均切割效率为46.23%。本研究拓宽了CRISPR/Cas9系统在作物中的使用范围,特别是NGA PAM位点较高的作物。     

Study on the Hepatoprotective Effect of Oxalis coriniculata L. and Related Mechanism by Regulating Oxidative Stress and TLR-2/NF-κB Signaling Pathway

[Objectives]This study aimed to explore the protective effect of Oxalis coriniculata L.on rats with acute liver injury induced by carbon tetrachloride(CCl4)and ...