水稻粒宽突变体gw87的基因定位及候选基因分析

【目的】对水稻粒宽突变体gw87（grain width 87）进行表型鉴定、遗传分析、基因定位及候选基因分析,为探明该基因调控水稻籽粒大小的分子机制及应用潜力奠定基础。【方法】利用甲基磺酸乙酯诱变籼稻恢复系材料676R,获得一份籽粒宽度和千粒重显著增加的突变体gw87。对该突变体进行表型观察、农艺性状调查及外源油菜素内酯（BL）敏感性、叶绿素含量、光合参数测定,了解其表型特征及生理特性。调查gw87与676R杂交F₁的表型和F₂群体的分离情况,分析其遗传行为;选取该群体中的突变植株进行高通量测序,并利用gw87与粳稻品种日本晴杂交的F₂代作为定位群体,通过MutMap分析和分子标记定位,遴选候选基因并进行DNA和cDNA测序验证。利用qRT-PCR分析gw87和676R中BR合成途径基因OsDWARF4、D11和D2的表达差异。【结果】与野生型亲本676R相比,gw87突变体的株高降低,节间缩短,其中,倒一节间长度缩短最大,并呈现出扭曲的形态;叶片长度减少,宽度增加;单株有效穗数、主穗穗长和结实率显著降低,但籽粒宽度和千粒重显著增大。BL敏感性试验显示,gw87突变体幼苗对外源BL的敏感性降低。光合色素和光合参数测定表明,gw87光合色素含量增加,光合速率也有所增加。遗传分析表明gw87的突变性状是由1对隐性核基因控制。MutMap分析显示gw87突变基因位于第5染色体中部,在该染色体区域仅有1个碱基突变引起编码氨基酸变化;分子标记连锁分析表明该突变基因位于InDel标记X2和X3之间约101 kb的染色体区域;综合这两方面分析结果,最后遴选出gw87候选基因是编码一个含有AP2/EREBP DNA绑定结构域的转录因子基因LOC_Os05g32270。对该候选基因进行DNA和cDNA测序验证,发现gw87突变体中该基因DNA的第1 041位的碱基由G突变为A,导致与该位点相邻的76 bp内含子序列被剪接为外显子,引起阅读框移码,蛋白翻译提前终止。qRT-PCR分析显示gw87突变体中BR合成途径基因的表达量显著上调,表明gw87突变体中BR信号减弱。【结论】gw87是smos1、shb、rla1、ngr5的新等位突变体,但与这些突变体表型不同,gw87籽粒的宽度和千粒重显著增加,可能是LOC_Os05g32270的突变位点不同,导致其编码蛋白的功能活性不同所造成。     

鸡脚重性状的全基因组关联分析

鸡(Gallus gallus)脚重性状为鸡产业中的重要经济性状.为了揭示鸡脚重性状的遗传基础,寻找可用于鸡脚改良的分子标记,本研究以核心群京海黄鸡为实验动物,测定其脚重,利用简化基因组测序技术在整个基因组范围内检测与脚重相关的SNPs.共检测到16个与脚重相关的SNPs位点,其中13个集中分布在4号染色体上72.8～77.9 Mb区域;2个SNP位点rs475641139和rs475548810达到5％ Bonferroni全基因组显著水平(P＜5.5E-07),其余位点达到全基因组潜在显著水平(P＜1.1E-05).筛选每个SNP周围1 Mb区域内的基因,共找到9个可能的候选基因,并使用基因本体论(Gene Ontology,GO)数据库对9个候选基因的细胞组分、分子功能和参与的生物学进程进行了分析.结果表明二氢蝶啶还原酶(dihydropteridine deductase,QDPR)基因、LIM域结合蛋白2(LIM domain-binding protein 2,LDB2)基因、类184B家族(family with sequence similarity 184 memberB,FAM184B)基因、复合信号免疫球蛋白J结合蛋白(recombination signal binding protein for immunoglobulin kappa J region,RBPJ)基因、纤维母细胞生长因子结合蛋白2(fibroblast growth factor-binding protein 2,FGFBP2)基因、富亮氨酸重复蛋白5(F-box and leucine-rich repeat protein 5,FBXL5)基因、胆囊收缩素受体A(cholecystokinin receptor type A,CCK1R)基因、肌动蛋白互作蛋白1 (WD repeat containing protein 1,WDR1)基因和类桶状蛋白4(tubby like protein 4,TULP4)9个基因和4号染色体72.8～77.9 Mb区域可能为影响鸡脚重性状的重要候选基因和潜在功能区域.本研究为鸡脚重性状的标记辅助选择提供了理论依据.     

陇东紫花苜蓿地上生物量及水分利用率分析

分析紫花苜蓿Medicago sativa地上生物量的变化表明:鲜、干质量曲线的增长特点是返青(萌芽)至现蕾后增长快,进入开花期后增长缓慢;第1茬鲜质量和干质量占全年总鲜质量和干质量的比例最大,高度与鲜质量和干质量的增长成正相关;在不同茬的生长期内,气象要素对其贡献不同,水分利用率以第2茬最高。     

褐飞虱自噬相关基因NlATG4的表达与功能

ATG4是一种半胱氨酸蛋白酶,负责ATG8蛋白羧基末端的切割,在自噬体形成过程中发挥重要作用。本研究克隆得到褐飞虱ATG4基因（NlATG4）的cDNA全长序列（GenBank登录号:MF062502.1）。RT-qPCR分析结果表明,NlATG4基因在褐飞虱的若虫和成虫时期均有表达,1～2龄若虫的表达量最高,在其他发育阶段的表达量稍低。RNAi结果显示,在dsRNA注射后的第4 d,靶标基因NlATG4的表达量显著下降（仅为dsGFP对照组的22%）,同时,NlATG4的RNA干扰导致虫体ATP含量显著降低,仅为1.0 μmol/(L·mg蛋白^-1),为对照组的25%;在注射后的第6 d,dsNlATG4处理组的褐飞虱存活率出现显著降低,其脂肪体组织变得松散,大型脂滴（直径≥5.0 μm）数量增多,类酵母共生菌周围泡状结构的界限基本消失;在注射后的第8 d,dsNlATG4处理的褐飞虱的存活率下降为36.7%,比dsGFP对照组的存活率下降53.3%。本研究表明靶向NlATG4基因的RNA干扰会破坏褐飞虱脂肪体的结构完整性和类酵母共生菌的微环境,影响脂类物质代谢和ATP含量,并导致褐飞虱存活率降低。因此,NlATG4基因在褐飞虱正常生长发育中发挥重要作用,在褐飞虱防治中具有一定的潜力。     

ULTRASONOGRAPHIC EVALUATION OF ADRENAL GLAND SIZE COMPARED TO BODY WEIGHT IN NORMAL DOGS

The accepted cut‐off value for adrenal gland maximum diameter of 0.74 cm to distinguish adrenal gland enlargement in dogs regardless of body weight may not be appropriate for small to medium breed dogs. The purpose of the current retrospective study was to examine adrenal gland dimensions as a function of body weight in healthy dogs in three weight categories (< 10 kg, 10–30 kg, and > 30 kg) representing small, medium, and large breeds, respectively, to establish greater confidence in determining if adrenal gland size is abnormal. The measurements of length (sagittal plane), cranial and caudal pole thickness (sagittal and transverse planes), and caudal pole width (transverse plane) of both adrenal glands were obtained ultrasonographically in clinically healthy dogs (n = 45) with 15 dogs in each weight group. Findings support our hypothesis that adrenal gland size correlates with body weight in normal dogs, and more precise reference intervals should be created for adrenal gland size by categorizing dogs as small, medium, or large breed. The caudal pole thickness of either adrenal gland in a sagittal plane was the best dimension for evaluating adrenal gland size based on low variability, ease, and reliability in measurement.     

Effects of sex and age on behavior and weight gain in beef calves after abrupt weaning

The effects of weaning on beef calves of different ages were investigated. Forty male and 40 female calves were either weaned at 6 (W6, n = 40) or 8 (W8, n = 40) months of age. The vocalization activity and behaviors (feeding, lying and standing/walking) were observed during the first 3 days following weaning. Body weight was recorded at 6 and 8 months and 2 weeks following weaning. W6 animals vocalized more often than W8 calves and females more than males on day 1 (P < 0.05). The vocalization activity decreased at a higher rate in W8 than in W6 from days 1 to 3. On day 2, W6 calves spent less time lying and more standing/walking than W8 calves (P < 0.05). The behaviors did not differ between males and females. The average daily gain after weaning did not differ between weaning ages (P > 0.05), but W8 calves gained more from 6 to 8 months of age. In conclusion, weaning distress was more pronounced in W6 than in W8 calves. W8 calves also had a superior growth performance during 6 to 8 months of age. The effect of the calves' sex was less pronounced. Therefore, it is recommended to wean beef calves later than 6 months of age.     

饲料脂肪源和乳化剂对牛蛙生长性能、肠道消化酶活力及肝脏生化指标的影响

本试验旨在研究以猪油或鱼油为脂肪源的饲料中添加乳化剂对牛蛙生长性能、肠道消化酶活力及肝脏生化指标的影响。试验采用2×2因子完全随机区组试验设计,2种脂肪源(猪油和鱼油)和2个乳化剂添加水平(0和300 mg/kg),配成4种等氮等能的试验饲料。将初始平均体重为(19.01±0.01)g的180只牛蛙随机分为4组,每组3个重复,每个重复15只蛙,进行为期8周的饲养试验。结果表明:未添加乳化剂的鱼油组增重率(WGR)和摄食量(FI)显著高于未添加乳化剂的猪油组(P0.05)。添加乳化剂的猪油组WGR和FI显著高于添加乳化剂的鱼油组(P0.05)。添加乳化剂的猪油组WGR、FI和特定生长率(SGR)显著高于未添加乳化剂的猪油组(P0.05),2组之间饲料系数(FCR)、氮沉积率(NRE)、能量沉积率(ERE)均差异不显著(P0.05)。添加乳化剂的鱼油组的各生长性能指标与未添加乳化剂的鱼油组均无显著差异(P0.05)。各组牛蛙胴体水分、粗蛋白质、粗脂肪和粗灰分含量差异不显著(P0.05),而添加乳化剂的猪油组全体粗脂肪含量显著高于未添加乳化剂的猪油组(P0.05)。各组牛蛙肠道蛋白酶和淀粉酶活力差异不显著(P0.05),饲料中添加乳化剂显著提高以猪油为主要脂肪源牛蛙的肠道脂肪酶活力(P0.05),而对以鱼油为主要脂肪源牛蛙的影响不显著(P0.05)。各组牛蛙肝脏过氧化氢酶(CAT)和超氧化物歧化酶(SOD)活力无显著差异(P0.05)。在以猪油为主要脂肪源时,添加乳化剂显著提高肝脏丙二醛(MDA)含量(P0.05);在以鱼油为主要脂肪源时,添加乳化剂对肝脏MDA含量无显著影响(P0.05)。根据结果得出,在以猪油为主要脂肪源的饲料中添加乳化剂可提高牛蛙的生长性能。     

基于全基因组填充重测序关联分析鉴别影响苏山猪初生体尺与乳头数性状的遗传位点

旨在鉴别影响苏山猪初生体尺和乳头数性状的遗传位点,开发可用于辅助育种的分子标记,为苏山猪生长和繁殖性能的持续选育提供理论基础。本试验对269头苏山猪初生仔猪(出生后24 h内)的体尺和乳头数表型进行测定,并采集耳组织样品。基于全基因组重测序及基因型填充策略,利用全基因组关联分析(genome-wide association study, GWAS)和群体分化指数(fixation index,Fst)的方法挖掘与目的性状强关联位点,并对位置功能候选基因开展GO和KEGG分析,确定最有可能的主效基因。本研究共鉴别到4个候选位点,分布在13、14和X染色体上,其中与初生体尺性状显著关联的位点有2个,与乳头数性状显著关联的位点有2个。本研究筛选出1个与体长性状相关的候选基因ACTA2;1个与胸围性状相关的候选基因COL4A6;3个与乳头数性状相关的候选基因ACTA2、CRTAP和SEPTIN6,为苏山猪初生体尺性状和乳头数性状的遗传改良提供了重要的分子标记。     

Characterization of an intravenous lipopolysaccharide inflammation model in calves with respect to the acute-phase response

Our objective was to develop a lipopolysaccharide (LPS) inflammation model in calves to evaluate the acute-phase response with respect to the release of pro-inflammatory cytokines and acute-phase proteins, fever development and sickness behaviour. Fourteen 4-week-old male Holstein Friesian calves were included and randomly assigned to a negative control group (n = 3) and an LPS-challenged group (n = 11). The latter received an intravenous bolus injection of 0.5 μg of LPS/kg body weight. Blood collection and clinical scoring were performed at 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 12, 18, 24, 28, 32, 48, 54 and 72 h post LPS administration (p.a.). In the LPS group, the following clinical signs were observed successively: tachypnoea (on average 18 min p.a.), decubitus (29 min p.a.), general depression (1.75 h p.a.), fever (5 h p.a.) and tachycardia (5 h p.a.). Subsequent to the recovery from respiratory distress, general depression was prominent, which deteriorated when fever increased. One animal did not survive LPS administration, whereas the other animals recovered on average within 6.1 h p.a. Moreover, the challenge significantly increased plasma concentrations of tumour necrosis factor-α, interleukin 6, serum amyloid A and haptoglobin, with peaking levels at 1, 3.5, 24 and 18 h p.a., respectively. The present LPS model was practical and reproducible, caused obvious clinical signs related to endotoxemia and a marked change in the studied inflammatory mediators, making it a suitable model to study the immunomodulatory properties of drugs in future research.     

荷斯坦奶牛牛嘴宽度影响因素分析及遗传参数估计

【目的】探究中国荷斯坦奶牛牛嘴宽度的群体规律,并估计其遗传参数。【方法】2021年7月至2021年8月分别在2个规模化奶牛场测定了628头泌乳期荷斯坦奶牛的牛嘴宽度及臀高、体高、体斜长和胸围等体尺性状,同时利用体斜长和胸围估计试验牛体重。利用SPSS 26.0软件分析了牧场、月龄和泌乳阶段等因素对牛嘴宽度的影响,以及牛嘴宽度与各体尺性状之间的相关关系;基于DMU软件采用单性状动物模型估计了牛嘴宽度的方差组分及遗传力。【结果】荷斯坦奶牛牛嘴宽度的群体平均值为17.28 cm,变异系数为5%,变异相对较小;牧场和月龄对牛嘴宽度均有极显著影响（P<0.01）,泌乳阶段对牛嘴宽度有显著影响（P<0.05）,牛嘴宽度随母牛月龄的增加呈逐渐增加的趋势,泌乳初期牛嘴宽度显著小于其他泌乳阶段（P<0.05）;牛嘴宽度与各体尺性状之间存在极显著正相关（P<0.01）;荷斯坦奶牛牛嘴宽度的遗传力估计值为0.28,为中高遗传力性状。【结论】牧场和月龄对奶牛牛嘴宽度影响极显著,泌乳阶段对奶牛牛嘴宽度影响显著,牛嘴宽度与臀高呈较弱相关。荷斯坦奶牛牛嘴宽度是一个具有中高遗传力的体尺性状。