羊草种子萌发相关基因的筛选及表达分析

前期实验表明,变温处理(28 ℃ 12 h/16 ℃ 12 h)可显著提高羊草种子的萌发率,且羊草种子萌发中的第1天是接受变温信号的关键时期。以此为研究基础,结合羊草种子变温萌发的转录组测序数据,针对羊草种子萌发初期对变温处理的响应筛选出与种子萌发、休眠及低温相关的基因24个,利用测序结果中这些基因的RPKM值制作基因表达热图并分析其表达差异。以萌发率高、低的两种羊草种质的种子为材料,对24个基因在恒温12 h(28 ℃)和变温1 d(28 ℃ 12 h/16 ℃ 12 h)萌发处理中的表达分别进行了定量分析。结果表明,与恒温对照相比,变温处理12 h后,表达明显上调的基因有SAIN1,PP2C62,EXPB3,EXPB4,GA3ox,EXPA2和EXPA7,而表达明显下调的基因有bHLH49,GID1,ABI8,Chi1,11833,CBF3,NAC2,PP2C72,SAIN2和5423。通过进一步分析相关基因在高、低萌发率两个种质中表达的差异,筛选出其中可能与羊草种子萌发相关的基因有几丁质酶基因Chi1,转录因子基因CBF3,羊草新基因5423,赤霉素合成基因GA3ox,细胞松弛素蛋白基因EXPB4和羊草新基因SAIN1,将为下一步阐明羊草种子萌发的分子作用机理奠定基础。     

Heritability and genetic correlations between rumination time and production traits in Holstein dairy cows during different lactation phases

So far, rumination has been used as a proxy for monitoring dairy cow health at farm level. However, investigating its genetic aspects as well as its correlation with other important productive traits may turn this management tool into a new informative selection criterion. However, scientific evidences on genetic correlation among rumination time (RT) and milk production and milk composition are still scarce. Therefore, the objective of this study was to estimate the heritability of RT across three lactation phases and its genetic correlation with milk production, milk composition and somatic cell count (SCC). Results of our study showed that heritability for RT was 0.34 and was constant across lactation. The mean genetic correlations between RT and milk production and composition traits were 0.07 (milk production), ?0.07 (protein yield), ?0.31 (fat yield), and ?0.32 (fat/protein ratio). The mean genetic correlation between RT and the SCC was 0.05.     

Bovine foetal sex determination—Different DNA extraction and amplification approaches for efficient livestock production

Foetal sex determination using polymerase chain reaction (PCR) in mammals is based on the amplification of gender‐specific foetal DNA sequences circulating in maternal blood. The bovine synepitheliochorial placenta does not allow a direct contact between the trophoblast and the maternal blood, resulting in difficult passage of foetal DNA and, consequently, its very small amounts in maternal bloodstream. Circulating cell‐free foetal DNA (ccffDNA) encompasses short nucleotide fragments (300–600 bp) in maternal circulation. The aim of this study was to assess this non‐invasive method in accurate prenatal sexing in early and late gestational periods in comparison with ultrasound diagnostics. As various DNA isolation and amplification methods were tested, their success in obtaining reliable results was evaluated. Two groups were tested, each consisting of 20 pregnant cows. Blood of a bull and a non‐pregnant heifer was the controls. Extraction of foetal DNA was accomplished by three different methods: using tubes with silicone membranes, a single‐tube extraction without silicone membranes and phenol–chloroform extraction. Following each extraction method, foetal DNA was amplified using PCR and real‐time PCR with both bAML and TSPY primers in a separate reaction. Positive results were obtained only after amplification of foetal DNA extracted with a single‐tube extraction kit. In comparison with ultrasound examination results and foetal gender recorded at birth, the sensitivity of the PCR test was 90% in Group I, but the technique failed to detect male foetuses in Group II. The real‐time PCR test sensitivity in Group I was 90% and in Group II 91.6%.     

Endocannabinoids as endometrial inflammatory markers in lactating Holstein cows

The objective of this study was to consider endocannabinoid system as inflammatory markers in bovine endometrium to better understand the role of this system in regulating many of the functions that are related to inflammatory condition. At day 26 post‐partum, fourteen cows were divided into two groups depending on the inflammatory condition: 1‐ subclinical endometritis (n = 7, with purulent or mucopurulent uterine discharge detectable in the vagina) and 2‐ healthy (n = 7, No (muco)) purulent discharge. Blood samples were collected at 26 and 30 days relative to calving to determine plasma tumour necrosis factor (TNF) and lipopolysaccharide‐binding protein (LBP) concentrations; moreover, uterine biopsy was carried out on day 26 post‐partum to measure mRNA abundance of TNF, interleukin‐1B (IL1B), interleukin‐6 (IL‐6), C‐X‐C motif chemokine ligand 8 (CXCL8), endocannabinoid receptor (CNR2), N‐acyl phosphatidylethanolamine phospholipase D (NAPEPLD), fatty acid amide hydrolase (FAAH), N‐acylethanolamine acid amidase (NAAA) and monoglyceride lipase (MGLL) by real‐time PCR. Results showed mean plasma concentrations of TNF and LBP were lower in healthy cows compared to subclinical endometritis cows (p < .05). Relative mRNA expression for NAAA and FAAH was decreased (p < .05), and relative mRNA expression for CNR2 and NAPEPLD increased in cows with subclinical endometritis compared to healthy cows. In conclusion, relative mRNA expression of TNF, IL1B and CXCL8 and plasma concentration of LBP increased during inflammatory condition along with decreased endocannabinoids hydrolyzing enzyme (NAAA and FAAH), increased enzymes that synthesize endocannabinoids (NAPEPLD) and relative gene expression of the endocannabinoid receptor; together, these contribute to increased endocannabinoids levels during inflammation. Overall, we provide evidence that endocannabinoid system is altered in endometrium tissue during inflammation through increased mRNA expression of CNR2 and synthesis enzyme and decreased mRNA expression of hydrolyzing enzymes interfere with pro‐cytokine production and signalling, which may interfere with the onset and progression of inflammation.     

不同种质玉米自交系种子萌发的干旱胁迫研究

种子萌发期被认为是作物生长的关键时期,也是衡量作物耐旱性强弱的重要时期。为了明确不同玉米自交系的耐旱性,本研究采用聚乙二醇(PEG)模拟干旱条件,对254份不同种质玉米自交系进行种子萌发期处理。通过检测发芽指数、发芽率、根数、根鲜重、根干重、茎鲜重、茎干重、根长、苗高等种子发芽性状,以其相对值为指标,鉴定不同自交系种子萌发期的耐旱性。结果表明,在5%PEG6000浓度胁迫时,种子萌发程度低,当PEG浓度为10%时,种子萌发受抑制程度较高；各性状相对指标都与品种综合抗旱能力呈显著或极显著正相关；不同自交系的耐旱性存在较大差异,通过隶属函数法结合抗旱分级标准筛选出99份中等耐旱材料。因此,该方法可以作为一种简单的玉米自交系萌发期耐旱性鉴定方法。     

阳春砂种子休眠与萌发过程中同工酶活性变化的研究

[目的]研究阳春砂种子休眠与萌发过程中同工酶的活性及酶谱变化规律,为阳春砂种子休眠与萌发过程生理调控机制的分析探讨奠定基础.[方法]以赤霉素浸种前、浸种后、培养10 d、种子露白、胚芽长至1 cm时各阶段的阳春砂种子为材料,测定其种子中过氧化物酶(POD)和超氧化物歧化酶(SOD)的活性,并采用聚丙烯酰胺凝胶电泳(PAGE)技术研究建立过氧化物酶(POD)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)及酯酶(EST)同工酶的电泳酶谱.[结果]阳春砂种子在休眠和萌发过程中,POD、SOD、CAT和EST同工酶的活性及酶谱带的数量均发生了一定的变化,其中POD和SOD同工酶活性均呈先减弱后增强的趋势;CAT同工酶在萌发前期变化不明显,后期酶的活性增强,且酶的种类也增加;EST同工酶的酶活性呈先减弱后增强再减弱的变化趋势,且在种子露白时活性达到最强.[结论]阳春砂种子休眠与萌发过程中同工酶谱的变化特点一定程度上可反应出种子休眠与萌发不同阶段的生理特性,这些同工酶基因的差异表达导致了阳春砂种子打破体眠和最终的萌发.     

穴盘规格对5种苔草属植物育苗的影响

穴盘育苗是苔草繁育的重要技术环节。为加快苔草的繁育速度,采用不同规格穴盘播种以研究青绿苔草(Carex breviculmis)、涝峪苔草(C.giraldiana)、披针苔草(C.lanceolata)、矮丛苔草(C.humilis var.nana)、脚苔草(C.pediformis)的适宜播种穴盘孔数对5种苔草出苗和幼苗生长的影响。通过对各种苔草的开始发芽时间、持续发芽时间、发芽率和幼苗的株高、叶数、单株鲜重等生长指标的分析,结果表明,6月温室穴盘播种育苗,青绿苔草、涝峪苔草穴盘育苗128孔或105孔穴盘较为适宜;披针苔草穴盘育苗宜采用105孔穴盘;矮丛苔草、脚苔草穴盘育苗较适宜选择288孔穴盘。     

温度对西藏5种野生豆科牧草种子萌发的影响研究

为探明毛果胡卢巴、劲直黄芪、天蓝苜蓿、镰荚棘豆和窄叶野豌豆5种野生豆科牧草种子萌发的适宜温度以提高其萌发率,研究了0/10℃ 、5/15℃ 、10/20℃ 、15/25℃ 、20/30℃ 、25/35℃ 、30/40℃等7种发芽温度对西藏5种野生豆科牧草种子萌发的影响.结果表明,5种野生豆科牧草种子在0/10℃条件下均无萌发;发芽率、发芽势、发芽指数和活力指数等指标随昼夜温度的升高呈先上升后下降趋势;毛果胡卢巴、劲直黄芪、天蓝苜蓿、镰荚棘豆和窄叶野豌豆牧草在适宜温度范围内种子萌发时滞最短,依次为2,2,2,3,4d;毛果胡卢巴和镰荚棘豆种子萌发最适温度为15/25℃,发芽率分别高达95%和812.5%;劲直黄芪、天蓝苜蓿、窄叶野豌豆种子萌发最适温度为20/30℃,发芽率依次为85%、912.5%和65%.     

PEG胁迫对西伯利亚冰草种子萌发及幼苗生理特性的影响

以PEG-6000模拟干旱胁迫,分析西伯利亚冰草种子发芽率、发芽势、胚根和胚芽长及三叶期幼苗相对电导率、丙二醛和游离脯氨酸含量的动态变化规律.结果表明:1)高渗透势PEG溶液(-0.2,-0.4 MPa)能够促进种子萌发,在-0.4 MPa时,发芽率和发芽势均达到峰值.2)高渗透势PEG溶液(-0.2,-0.4,-0.6 MPa)对胚根伸长具有一定的促进作用,-0.4 MPa最大,但对胚芽表现出一定的抑制作用.3)随着渗透势的降低,发芽率、发芽势、胚根长和胚芽长不断下降,胚根/胚芽的比值持续上升.4)3叶期幼苗相对电导率、丙二醛和脯氨酸含量随胁迫强度的增加,胁迫时间的延长持续增加,相对电导率、丙二醛含量除了-0.4 MPa,1d处理外,其他处理均极显著(p＜0.01)高于对照,脯氨酸含量各处理均极显著(p＜0.01)高于对照,相对电导率、丙二醛和脯氨酸这3个指标均可作为抗旱性评价指标.