Multimarker and rare variants genomewide association studies for bone weight in Simmental cattle

Bone weight, defined as the total weight of the bones in all the forequarter and hindquarter joints, can reflect somebody conformation traits and skeletal diseases. To gain a better understanding of the genetic determinants of bone weight, we used a composite strategy including multimarker and rare‐marker association to perform genomewide association studies (GWAS) for that character in Simmental cattle. Our strategy consisted of three models: (i) A traditional linear mixed model (LMM) was applied (Q+K‐LMM); (ii) single nucleotide polymorphisms (SNPs) with p‐values less than .05 from the LMM were selected to undergo the least absolute shrinkage and selector operator (Lasso) in the second stage (LMM‐Lasso); (iii) genes containing two or more rare SNPs were examined by performing the sequence kernel association test (gene‐based SKAT). A total of 1,225 cattle were genotyped with an Illumina BovineHD BeadChip containing 770,000 SNPs. After the quality‐control procedures, 1,217 individuals with 608,696 common SNPs and 105,787 rare SNPs (with 0.001 < minor allele frequency [MAF] <0.05) remained in the sample for analysis. A traditional LMM successfully mapped three genes associated with bone weight, while LMM‐Lasso identified nine genes, which included all genes found by traditional LMM. Only a single gene, EPHB3, surpassed the significance threshold after Bonferroni correction in gene‐based SKAT. In conclusion, based on functional annotation and results from previous endeavours, we believe that LCORL, RIMS2, LAP3, PRKAR2B, CHSY1, MAP2K6 and EPHB3 are candidate genes for bone weight. In general, such a comprehensive strategy for GWAS may be useful for researchers seeking to probe the full genetic architecture underlying economic traits in livestock.     

Disaccharide combinations and the expression of enolase3 and plasma membrane Ca2+ ATPase isoform in sturgeon sperm cryopreservation

Acipenser sinensis and Acipenser dabryanus are critically endangered species, so germplasm conservation via cryopreservation of sperm is necessary. Disaccharides can act as membrane‐impermeable cryoprotectants, and enolase3 (ENO3) and plasma membrane Ca²⁺ ATPase isoform (PMCA2) are proteins associated with sperm quality. We considered seven characteristics of sperm quality in cultured brood stock from A. sinensis and A. dabryanus. We tested use of sucrose or trehalose alone and in combination at different concentrations for cryopreservation of A. dabryanus sperm. A low concentration of sucrose plus trehalose (S₁₅T₁₅) was optimal. Mixing of the extender with sucrose, lactose, or trehalose alone or with pairwise mixtures revealed that a mixture of lactose and trehalose (L₁₅T₁₅) gave the best results for both A. sinensis and A. dabryanus. Enolase3 and PMCA2 expression levels were measured in cryopreserved A. sinensis sperm via Western blotting. Relative ENO3 and PMCA2 expression levels were examined, and the relationship between disaccharide composition, sperm quality and protein expression was explored in A. sinensis. The results showed that relative ENO3 and PMCA2 expression levels were the highest at L₁₅T₁₅ in cryopreserved A. sinensis sperm. There were significant positive correlations between ENO3 expression and percentage membrane integrity, and between PMCA2 expression and sperm motility parameters (percentage of motile sperm, curvilinear velocity, straight‐line velocity and average path velocity; p < .05) in cryopreserved A. sinensis sperm. Our results indicate the optimal disaccharide combination and concentrations for cryopreservation of A. sinensis and A. dabryanus sperm and suggest that ENO3 and PMCA2 expression levels could serve as a valuable indicator of sperm quality in A. sinensis.     

Effects of five cryoprotectants on proliferation and differentiation‐related gene expression of frozen‐thawed bovine calf testicular tissue

The cryopreservation of testicular tissue is a potential method for preserving male fertility. However, the effect of cryopreservation on bovine calf testicular tissue is scarce. This study investigated the effect of different cryoprotectants on bovine calf testicular tissue at the molecular level. Testicular tissue from ten immature bovine calves (6 months) was collected after slaughter and cryopreserved in an extender containing different concentrations of the following five cryopreservation solutions (CP): bovine serum albumin (BSA) with 5% dimethyl sulfoxide (DMSO), trehalose with 5% DMSO, DMSO and glycerol and ethylene glycol (EG). After 7‐day cryopreservation, the expression levels of three spermatogonial stem cell (SSC)‐related genes, octamer‐4 (OCT4), KIT ligand (MGF/SCF) and kit oncogene (C‐KIT), were investigated by quantitative PCR (qPCR). The cell viability was highest for the tissues preserved with 30 mg/ml BSA (77.82% ± 1.22) and 40 mg/ml trehalose (74.23% ± 1.16) compared with other groups (p < 0.05), and the level of expression of the three genes was highest with 30 mg/ml BSA (p < 0.05). Compared with other CPs, the 30 mg/ml BSA and 40 mg/ml trehalose have the better cryopreserve protection. The 30 mg/ml BSA is the most viable media for the cryopreservation of testicular tissue from cattle.     

紫花苜蓿茎秆组织中木质素的分布与沉积模式

以甘农5号紫花苜蓿(Medicago sativa‘Gannong No.5’)为材料,采用组织化学染色方法研究了茎秆中木质素的发生、分布与沉积规律。结果表明,苜蓿茎秆中木质素的分布与生长部位密切相关,在茎秆顶端初生维管组织中仅木质部有木质素分布,随节间下移,木质素开始在初生韧皮纤维、次生木质部和髓射线中大量沉积;茎秆中存在G、S两种木质素,S木质素的发生迟于G木质素;在维管束之间,两种木质素均以"厚角组织处维管束→厚角组织间维管束"的模式沉积,但在维管束内,木质素沉积表现出异质性,G木质素沉积模式为"初生木质部导管→初生韧皮纤维→次生木质部和髓射线";S木质素沉积模式为"初生韧皮纤维→次生木质部和髓射线→髓薄壁细胞"。分析认为,紫花苜蓿茎秆中木质素特殊的沉积模式可能是其对北温带生长环境的一种适应对策。     

紫茎泽兰对萨能奶山羊血常规及重要脏器病理学的影响

紫茎泽兰(Eupatorium adenophorum)在中国作为入侵物种,可造成食草动物中毒死亡,并给畜牧业带巨大的损失。为研究反刍动物采食紫茎泽兰后的中毒机理,本研究探讨了日粮中添加不同剂量的紫茎泽兰对萨能奶山羊(Saanen goat)血常规及肝、脾、肾的病理学影响。选择16只5~6月龄健康萨能奶山羊,随机分成对照组(日粮中未添加紫茎泽兰)和试验Ⅰ、Ⅱ、Ⅲ组(日粮配比中分别添加紫茎泽兰40%、60%和80%),每组4只。试验Ⅰ、Ⅱ、Ⅲ组分别饲喂含40%、60%和80%紫茎泽兰的混合日粮3个月,每两周检测一次血常规,试验末屠宰奶山羊,并观察其肝、肾、脾的病理学变化。结果表明,与对照组相比,各试验组血液中的白细胞(white blood cell,)和中性粒细胞(neutrophil,NEUT)均显著升高(P0.05),试验后期极显著高于对照组(P0.01);而各试验组血液中淋巴细胞(lymphocyte,LY)与血红蛋白(hemoglobin,HGB)均呈下降趋势,试验后期均极显著低于对照组(P0.01)。肝和脾肿大,肝组织出现淤血,肝细胞发生水泡变性及脂肪变性等,肾组织伴有出血、坏死、颗粒变性及水泡样变性的现象。因此,本研究结果说明紫茎泽兰对萨能奶山羊血液指标有显著影响,且长期摄入紫茎泽兰可引起动物肝、肾、脾等主要实质器官不同程度的损伤。     

基于冗余分析的高寒草原土壤与草地退化关系

在高寒草原退化草地设置研究样地,观测植物群落特征、采集土壤样品、分析土壤物理化学性质,依据数量生态学基本原理,探讨高寒草原退化草地与土壤因子间关系。结果表明:不同退化程度样地沿冗余分析排序图第一排序轴分布,第一排序轴反映草地退化程度的变化;草地植物群落中与第一排序轴负相关且按相关程度大小排序的指标为盖度>地上生物量>地下生物量;第一、第二排序轴能够解释97.1%的土壤因子与草地退化关系;土壤沙砾、p H与第一排序轴正相关,土壤有机碳、土壤含水量、容重、全氮、有效氮与第一排序轴负相关;第一排序轴及所有排序轴所反映的土壤因子均与草地退化之间呈极显著相关关系(P<0.01);不同土壤因子与草地退化之间相关程度不同,土壤有机碳(r=-0.890)、土壤含水量(r=-0.864)、容重(r=-0.847)、全氮(r=-0.836)、有效氮(r=-0.821)等与高寒草原退化相关程度更高且相关关系极显著(P<0.01)。利用退化草地群落特征和土壤因子数据矩阵进行冗余分析能够较好地综合反映土壤因子与草地退化之间的关系及相关程度,土壤有机碳、土壤含水量、容重、全氮、有效氮是反映高寒草原退化的重要土壤指标。     

近15年新疆伊犁河谷草地退化时空变化特征

以伊犁河谷为研究区,利用MODIS NDVI数据及像元二分模型,反演草地植被覆盖度,以草地植被覆盖度为评价标准,结合数字高程模型(digital elevation model,DEM)数据及Getis-Ord Gi*冷/热点分析方法,对伊犁河谷2001-2015年草地退化的时空特征进行了分析。结果表明,1)受持续过度放牧及气候条件影响,2001-2015年伊犁河谷草地整体持续退化,15年内退化草地比例达46.18%,但退化以轻度为主;2)空间上退化草地的分布范围逐步向高海拔区域扩展,海拔1 500-3 000 m的中山和中高山区退化草地扩张最明显;3)草地生态保护政策的实施减缓了草地退化速度,草地退化与改善的空间差异逐渐明显,以退化为主的单一变化趋势有所改变;4)利用NDVI反演植被覆盖度对草地退化进行评价的方法存在对高植被覆盖区域草地退化敏感性相对较弱的缺陷。     

基质对黑果腺肋花楸幼苗生长的影响

以黑果腺肋花楸(Aronia melanocarpa)为试验材料,选用田园土、泥炭土、珍珠岩和黄沙,采用正交试验设计,研究不同基质配比对黑果腺肋花楸成苗率以及幼苗形态指标和生理指标的影响。结果表明,不同育苗基质对黑果腺肋花楸出苗率、苗高、地径、主根长、侧根数和高径比等形态指标以及根系活力、可溶性糖、淀粉和蛋白质等生理指标的影响差异显著,综合分析各项指标,筛选出适合黑果腺肋花楸容器育苗的栽培基质为田园土(40%)+泥炭土(30%)+黄沙(30%)、田园土(40%)+珍珠岩(30%)+黄沙(30%)、田园土(40%)+泥炭土(30%)+珍珠岩(30%)和田园土(60%)+泥炭土(20%)+黄沙(20%)。