玉米高密度SNP遗传图谱构建及其秃尖QTL定位

[目的]构建在秃尖性状上存在明显差异的玉米高密度SNP遗传图谱,并对其秃尖QTL进行定位,为玉米秃尖分子机理研究及玉米抗秃尖品种选育提供理论参考.[方法]以无秃尖性状的自交系S群411331为母本、有秃尖性状的自交系综53313为父本,通过杂交和自交获得F2代群体.利用容量达10K的分子芯片获取大量SNP分子标记,从中筛选出具有多态性的SNP分子标记,构建F2代群体的高密度遗传图谱,并结合秃尖表型数据,分别采用QTL定位软件Rqtl和QTL.gCIMapping.GUI 1.1对相应的秃尖QTL位点进行鉴定及定位.[结果]F2代群体的平均秃尖长度为4.25 cm,说明其秃尖性状更偏向于父本综53313,秃尖整体较长,且秃尖变幅为0~6.1 cm,偏度和峰度值均位于-1.00~1.00,符合数量性状的分布特征.从2612个多态SNP分子标记中共筛选出2599个SNP分子标记成功构建遗传连锁图谱,总图距5624.38 cM,标记间平均距离2.27 cM.利用Rqtl共检测到6个QTL,分别位于第3、4、5、6、8和9染色体,其中加性效应和显性效应最大的2个QTL分别位于第6和8染色体,解释遗传变异的14.4%和16.3%.利用QTL.gCIMapping.GUI 1.1共检测到9个QTL,分别位于第1、4、5、6、8和9染色体,显性效应和加性效应最大的2个QTL分别位于第6和8染色体,与Rqtl检测结果相比,二者均在第8和9染色体的同一位置检测到1个QTL,在第5染色体检测到的QTL位置也较邻近,且效应最大的2个QTL均位于第6和8染色体;不同之处在于Rqtl在各染色体上只检测到1个QTL,而QTL.gCI-Mapping.GUI 1.1在第6和8染色体分别检测到2和3个QTL,在第1染色体检测到1个QTL,在第3染色体未检测到QTL,而Rqtl检测出的第1和3染色体QTL情况相反.[结论]玉米秃尖QTL分别位于第1、3、4、5、6、8和9染色体,其中主效QTL在第6和8染色体.     

OCX-32基因内含子变异对长顺绿壳蛋鸡蛋壳品质的影响

[目的]探讨OCX-32基因对长顺绿壳蛋鸡蛋壳品质的遗传效应,为长顺绿壳蛋鸡的保种选育及开发利用提供参考依据.[方法]以长顺绿壳蛋鸡为材料,采用PCR产物直接测序法筛选OCX-32基因SNP多态位点,实时荧光定量PCR检测组织表达谱,运用SPSS 19.0广义线性模型(GLM)分析SNP位点基因型或双倍型与所测定性状指标的相关性.[结果]在长顺绿壳蛋鸡OCX-32基因中检测到3个SNPs位点,分别是位于内含子1上的g.22592323G>T及内含子3上的g.22597350T>C和g.22597555G>A.卡方(χ2)检测结果发现,g.22597350T>C位点的基因型分布显著偏离Hardy-Weinberg平衡(P<0.05,下同).连锁不平衡分析结果显示,3个SNPs突变位点不存在强连锁不平衡,共发现4种单倍型和8种双倍型,单倍型H1和双倍型H1H2频率最高,分别为0.523和0.345.实时荧光定量PCR检测结果显示,OCX-32基因在长顺绿壳蛋鸡12个组织中存在不同程度的表达,表达量排序为肾脏>心脏>子宫>腹脂>小肠>脾脏>肝脏>腺胃>胸肌>胰腺>肺脏>肌胃.关联分析结果显示,g.22597350T>C位点CC基因型在蛋壳强度和蛋壳重2个品质指标上显著高于CT基因型和TT基因型;双倍型H4H4个体的蛋壳强度显著高于其他7种双倍型个体,蛋壳重显著高于H2H4个体.[结论]OCX-32基因内含子变异能影响长顺绿壳蛋鸡蛋壳品质,检测到的3个SNPs位点可作为鸡蛋壳品质选择的遗传分子标记,其中突变位点g.22597350T>C的CC基因型和双倍型H4H4是影响蛋壳强度和蛋壳重的关键基因型和双倍型,有利改善蛋壳品质.     

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.     

Long-term fertilization leads to specific PLFA finger-prints in Chinese Hapludults soil

Soil microbes play essential roles in the biogeochemical processes of organic carbon and nutrient cycling. Many studies have reported various short-term effects of fertilization on soil microbes. However, less is known about the effects of longterm fertilization regimes on the rhizosphere. Therefore, the objective of this study was to explore how the soil microbial communities in the rhizosphere respond to different long-term fertilization strategies. Based on a 21-year field treatment experiment in Guizhou, China, we extracted phospholipid fatty acids(PLFAs) to determine the microbial community structure in both the non-rhizosphere(NR) and rhizosphere(R). Six treatments were included: no fertilizer(CK), mineral nitrogen fertilizer(N), N with potassium(NK), phosphorus with K(PK), NPK, and NPK combined with manure(MNPK). The results showed that total PLFAs under unbalanced mineral fertilization(N, NK and PK) were decreased by 45% on average in the NR compared with CK, whereas MNPK increased fungi and G~– bacteria abundance significantly in both the NR(by 33 and 23%) and R(by 15 and 20%), respectively. In addition, all microbial groups in the R under these treatments(N, NK and PK) were significantly increased relative to those in the NR, except for the ratio of F/B and G~+/G~–, which might be due to the high nutrient availability in the R. Soil pH and SOC significantly regulated the soil microbial community and structure, explaining 51 and 20% of the variation in the NR, respectively. However, the rhizosphere microbial community structure was only significantly affected by soil pH(31%). We concluded that the soil microbial community in the NR was more strongly affected by long-term fertilization than that in the R due to the rhizosphere effect in the agricultural ecosystem. Rhizosphere nutrient conditions and buffering capacity could help microbial communities resist the change from the long-term fertilization.     

间作香露兜提高槟榔根系生长和土壤酶活性

采用盆栽试验的方法,对槟榔单作、香露兜单作和槟榔间作香露兜3种栽培模式下根系生长发育和土壤酶活性进行比较研究。结果表明：槟榔间作香露兜促进了槟榔干物质累积,且分配到根系的干物质相对增多,地下部干物质质量显著增加,对香露兜的影响不显著;间作后显著增加槟榔叶片SPAD值,对香露兜无显著影响;间作模式下槟榔总根长、根系总表面积、根数目显著增多,分别比单作增加了78.64%、50.96%、81.22%,而且间作对槟榔根系生长发育的促进作用大于香露兜;槟榔间作香露兜后,土壤酸性磷酸酶、过氧化氢酶和过氧化物酶活性均显著高于槟榔单作,土壤脲酶与槟榔单作无显著差异,但显著低于香露兜单作;土壤过氧化物酶活性与根数目和根体积呈显著正相关。综上所述,槟榔与香露兜根系之间竞争较小,与槟榔单作相比,槟榔间作香露兜模式能够促进槟榔根系生长,提高土壤酶活性。     

茶园土壤-茶树-茶叶-茶汤中镉转移规律及安全性评价

为论证镉污染土壤种植茶树的安全性与可行性,通过盆钵茶园试验研究了湖南9个主栽茶树品种在不同镉含量土壤中对镉的吸收积累特性和茶汤中镉元素的浸出率。结果表明,在pH值4.1的土壤中,通过根系吸收的镉元素从根系到主干,再到侧枝枝干和茶树叶片的转运过程中,各器官的镉含量依次递减,茶树嫩芽(叶)中镉的富集系数仅0.13~0.24;茶树种植在土壤镉含量低于3.0 mg/kg的土壤中,茶叶没有出现镉超标现象;镉元素在茶汤中的平均浸出率为9.87%,利用膳食暴露风险评估模型对茶叶和茶汤中的镉含量进行风险评估,表明供试茶叶样品中的镉不会对人体健康产生威胁。     

Minimally modified low-density lipoprotein up-regulates endothelin receptors in mouse mesenteric artery through p38 MAPK pathway

AIMTo investigate whether minimally modified low-density lipoprotein (mmLDL) affects the quantity and activity of endothelin （ET） type A (ET_A) and type B (ET_B) receptors in mouse mesenteric artery by activating p38 mitogen-activated protein kinase (MAPK) inflammatory pathway. METHODSThe KM mice were divided into normal saline (NS) group (injection of NS via caudal vein), mmLDL group (injection of mmLDL via caudal vein), LDL group (injection of LDL via caudal vein), mmLDL+SB 203580 group (injection of mmLDL via caudal vein and intraperitoneal injection of p38 MAPK pathway specific inhibitor SB 203580) and mmLDL+DMSO group (injection of mmLDL via caudal vein and intraperitoneal injection of DMSO). Mesenteric artery ring segment vasoconstriction dose-response curves affected by sarafotoxin 6c (S6c) and ET-1 were recorded by the myography system. The mRNA levels of ET_B receptor, ET_A receptor and interleukin-6 (IL-6) were detected by RT-qPCR. The protein levels of ET_B receptor, ET_A receptor, IL-6, p38 MAPK, p-p38 MAPK, NF-κB and p-NF-κB were determined by Western blot. The serum concentration of IL-6 was measured by ELISA. RESULTSThe contractile responses of the blood vessel segments to S6c and ET-1 were significantly increased by mmLDL (P<0.01). The mRNA and protein expression levels of ET_A receptor, ET_B receptor, and IL-6 significantly increased (P<0.01). The protein levels of p-p38 MAPK and p-NF-κB were significantly increased (P<0.01). The serum level of IL-6 was significantly increased (P<0.01). These effects of mmLDL were inhibited by p38 MAPK inhibitor SB 203580. CONCLUSION mmLDL increses the serum concentration of IL-6, up-regulates the expression of IL-6, ET_A receptor and ET_B receptor in mouse mesenteric artery, and enhances the vasoconstriction function medi?ated by ET_A and ET_B receptors, which is related to the activation of p38 MAPK inflammatory pathway and downstream NF-κB pathway.     

Roles of protein phosphatase 4 in down-regulation of eNOS Ser633 phosphorylation induced by palmitic acid in human umbilical vein endotheli?al cells

AIMTo investigate the roles of protein phosphatase 4 (PP4) in down-regulation of endothelial nitric oxide synthase (eNOS) Ser633 phosphorylation induced by palmitic acid (PA). METHODSHuman umbilical vein endothelial cells (HUVECs) were treated with PA at 25 μmol/L, 50 μmol/L, 100 μmol/L and 200μmol/L for 36 h, or treated with PA at 100 μmol/L for 12 h, 24 h, 36 h and 48 h. Protein phosphatase 2A (PP2A) family inhibitor fostriecin (FST, 20 nmol/L) or okadaic acid (OA, 5 nmol/L) was selected to pretreat the HUVECs for 30 min. Protein phosphatase 4 catalytic subunit (PP4c) siRNA or protein phosphatase 2A catalytic subunit (PP2Ac) siRNA was transfected into the HUVECs. The protein expression levels of of eNOS, PP4c and PP2Ac, as well as the level of eNOS Ser633 phosphorylation, were detected by Western blot. The intracellular nitric oxide (NO) content was measured by DAF-FM DA. RESULTS(1) Compared with control group, the levels of eNOS Ser633 phosphorylation were decreased in PA groups in which the HUVECs were treated with 25 μmol/L, 50 μmol/L, 100 μmol/L and 200 μmol/L PA for 36 h (P<0.05) and 100 μmol/L PA for 24 h, 36 h and 48 h (P<0.05). No significant difference in the level of total eNOS protein expression among all the groups was observed. (2) Compared with control group, both FST and OA pretreatment reversed the reduction of eNOS Ser633 phosphorylation (P<0.05) and the decrease in intracellular NO content (P<0.05) induced by PA. No significant difference in the level of total eNOS protein expression among all the groups was observed. (3) Compared with si-Control group, the PP4c protein expression was significantly reduced (P<0.05), while the level of eNOS Ser633 phosphorylation was significantly increased in si-PP4c group (P<0.05). Although the levels of PP2Ac protein expression declined significantly (P<0.05), the level of eNOS Ser633 phosphorylation remained unchanged in si-PP2Ac group. No significant differencein the level of total eNOS protein expression among all the groups was found. CONCLUSION PA significantly reduces the level of eNOS Ser633 phosphorylation and the content of NO in the HUVECs, which may be due to PA inducing the activation of the PP2A family member PP4 rather than PP2A.     

‘越心’草莓组培突变体着色差异的分子机理初探

初步探讨了在‘越心’草莓（Fragaria × ananassa‘Yuexin’）茎尖离体培养再生植株中发现的稳定变异突变体着色差异的分子机理。分析结果显示,突变体与‘越心’在植株形态、始花期、始果期、始熟期、单果质量、平均株产、果形、风味等方面均无明显差异;而突变体外果皮色泽较淡,L*值较高,a*值较低,H色度角值较大,颜色指数CIRG显著小,且果肉不着色（‘越心’果肉红色）。突变体果实总花青苷含量为‘越心’的13.2%,其中以天竺葵素–3–O–葡萄糖苷（Pg3G）含量差异最大,突变体Pg3G含量仅为‘越心’的12.9%。花青苷合成途径结构基因FaF3H、FaCHS、FaDFR、FaANS、FaCHI、Fa3GT的表达水平在突变体中显著降低,这可能是突变体成熟果实花青苷水平显著低于‘越心’的直接原因。FaMYB10和FaMYB1在突变体中的表达水平显著下降,且FaMYB10在几个主成分中的特征向量绝对值均显著较高,进一步证实了MYB10是草莓成熟过程中花青苷合成的类黄酮或苯丙烷类化合物途径主要调节因子之一。提取叶片基因组DNA进行基于KASP标记的基因分型结果显示,74对KASP引物在‘越心’草莓及其突变体中的基因分型不存在差异。本结果初步证实突变导致参与调控花青苷合成途径的关键基因表达发生显著下调从而使花青苷积累显著减少,果实着色变淡。     

草地贪夜蛾对我国玉米产业的潜在经济损失评估

草地贪夜蛾Spodoptera frugiperda,又称秋黏虫,起源于美洲。自2019年1月入侵我国云南以来,目前已扩散至我国25个省、市(区)。目前在我国发现的草地贪夜蛾主要为害玉米,本研究在收集了草地贪夜蛾的为害及防治情况,玉米的产量、面积、价格等相关数据的基础上,利用随机模型@RISK分别预测了其在防治与不防治场景下对我国玉米产业的潜在经济损失。结果表明,在不防治场景下,草地贪夜蛾对我国玉米的潜在经济损失总量的90%置信区间为375.68亿～3 283.45亿元,投入防治后可挽回的潜在经济损失90%置信区间为254.78亿～2 918.93亿元。因此,为保护我国玉米种植业的安全生产,应加强田间防治来降低其对我国玉米产业造成的潜在经济损失。