刺萼龙葵种子中适宜内参基因的筛选

入侵杂草刺萼龙葵Solanum rostratum Dunal传播扩散的主要载体是种子,研究其种子休眠萌发基因的激素调控对于其防除具有重要意义,而选择合适的内参基因可以提高相关基因表达分析的准确性。本研究以赤霉素、脱落酸和水处理的刺萼龙葵种子为材料,利用geNorm、NormFinder、BestKeeper和RefFinder 4种软件对15个候选内参基因进行表达稳定性评价,并通过检测ABI5(abscisic acid-insensitive 5)的表达验证所筛选的内参基因的适用性。结果表明,对于赤霉素、脱落酸和水处理过的种子,最稳定的内参基因分别为eIF(eukaryotic initiation factor)、SAND(SAND protein family)和ACT(β-actin);对所有种子样本而言,PP2Acs(a catalytic subunit of protein phosphatase 2A)是最稳定的内参基因。研究结果将为刺萼龙葵种子休眠萌发的遗传调控研究提供重要参考。     

利用凝胶扩散法测定刺萼龙葵种子中β-甘露聚糖酶的活性

β-甘露聚糖酶是种子萌发过程中降解胚乳细胞壁的关键酶,明确其活性的动态变化可为揭示杂草种子的休眠萌发机制提供重要依据。以外来杂草刺萼龙葵Solanum rostratum Dunal种子为材料,建立了种子中β-甘露聚糖酶活性的检测方法—凝胶扩散法。利用凝胶扩散法对不同贮存时间及贮存条件下刺萼龙葵种子中β-甘露聚糖酶的活性进行了检测,发现贮存3年以上的种子中该酶的活性为0.03 nmol/(min·mg),显著低于贮存3年以下的种子中的酶活性0.15 nmol/(min·mg),而湿润冷藏的种子中β-甘露聚糖酶的活性为0.12 nmol/(min·mg),显著高于干燥冷藏的种子的酶活性0.02 nmol/(min·mg)。实际应用结果表明,凝胶扩散法综合了传统方法的优势,检测特异性强、灵敏度高、重复性好,可同时检测大量种子样品,具有良好的实用性。     

Temporal and spatial progress of the diseases caused by the crinivirus tomato chlorosis virus and the begomovirus tomato severe rugose virus in tomatoes in Brazil

Efficient management of whitefly-borne diseases remains a challenge due to the lack of a comprehensive understanding of their epidemiology, particularly of the diseases tomato golden mosaic and tomato yellowing. Here, by monitoring 16 plots in four commercial fields, the temporal and spatial distribution of these two diseases were studied in tomato fields in Brazil. In the experimental plots these diseases were caused by tomato severe rugose virus (ToSRV) and tomato chlorosis virus (ToCV), respectively. The incidence of each virus was similar in the plots within a field but varied greatly among fields. Plants with symptoms for both diseases were randomly distributed in three of four spatial analyses. The curves representing the progress of both diseases were similar and contained small fluctuations, indicating that the spread of both viruses was similar under field conditions. In transmission experiments of ToSRV and ToCV by Bemisia tabaci MEAM1 (former biotype B), these viruses had a similar transmission rate in single or mixed infections. It was then shown that primary and secondary spread of ToCV were not efficiently controlled by insecticide applications. Finally, in a typical monomolecular model of disease progress, simulation of the primary dissemination of ToSRV and ToCV showed that infected plants were predominantly randomly distributed. It is concluded that, although the manner of vector transmission differs between ToSRV (persistent) and ToCV (semipersistent), the main dispersal mechanisms are most probably similar for these two diseases: primary spread is the predominant mechanism, and epidemics of these diseases have been caused by several influxes of viruliferous whiteflies.     

番茄褪绿病毒(tomato chlorosis virus)的新寄主:水茄(Solanum torvum Swartz)

Tomato chlorosis virus (ToCV) is a plant virus that is mainly propagated by Bemisia tabaci in a semi-persistent and non-circulative manner.It has a wide range of host plants,and has been reported in many countries,causing serious economic losses in vegetable production.In 2019,we investigated about 10 fields,one ha each in Shouguang,Shandong province (China),and in each field we observed symptoms of interveinal chlorosis on lower leaves of the Solanum torvum Swartz,and a large number of B.tabaci gathered on the back of its leaves.To determine the presence of ToCV,total RNA of S.torvum was extracted followed by RT-PCR.The 1 074 (GenBank accessions number MN545620) and 466 bp (GenBank accessions number MN545621) fragments were gel purified and sequenced.The sequences shared 99.44% and 99.57% similarity with ToCV reference sequence tomato chlorosis virus segment RNA1 (AY903447) and RNA2 (AY903448).The results of insect transmission test confirmed that ToCV can spread from S. torvum to tomato.This study confirms S.torvum as a newly reported host of ToCV.     

赤霉素不同处理浓度和浸种时间对托鲁巴姆发芽的影响

托鲁巴姆种子的休眠较深,发芽较困难。为了打破其种子的休眠期,加快种子萌发,提高种子发芽的整齐度,以托鲁巴姆为试材,研究了不同的赤霉素处理浓度和浸种时间对其发芽的影响。结果表明:生产中适宜的赤霉素处理浓度为750~1 250 mg/L,浸种时间为36 h,发芽率可达93.00%;可有效提高托鲁巴姆种子的发芽速率和发芽整齐度。     

基于侯选基因标记的四倍体马铃薯休眠QTL关联分析

休眠期是马铃薯(SolanumtuberosumL.)重要的块茎性状之一,寻找调控马铃薯块茎休眠的关键基因,揭示其分子机制以选育具有适宜休眠期长度的马铃薯品种,对于解决当前马铃薯产业中过长或过短休眠期带来的经济损失和食品安全隐患等问题十分关键。前期研究在二倍体马铃薯连锁群体中定位了6个加性休眠QTL,本研究拟在四倍体马铃薯育种材料中验证这些休眠QTL。基于休眠QTL连锁的候选基因标记,采用混合线性模型(MLM),模型中考虑群体结构和亲缘关系(Q+K),在四倍体马铃薯自然群体St-hzau中对马铃薯块茎休眠期进行了关联分析。5号染色体上休眠QTL DorB5.3连锁的候选基因标记S199_300和GWD (根据葡聚糖水双激酶α-glucan water dikinase基因设计)与马铃薯块茎休眠期具有显著的关联(P0.05),分别解释了休眠期表型变异的7.8%和3.2%,分别能增加休眠期7.1 d和4.5 d,即在二倍体马铃薯连锁群体中定位的稳定主效休眠QTL DorB5.3在四倍体马铃薯关联群体St-hzau中也表现显著, DorB5.3的稳定性在关联分析结果中得到了验证,表明候选基因标记策略在马铃薯块茎休眠QTL关联分析中是一种有效的策略。本研究所验证的主效休眠QTL DorB5.3及相应连锁标记可以直接用于马铃薯休眠育种。据此可以推测GWD可能在控制还原糖含量和块茎休眠2个方面均发挥作用,马铃薯块茎休眠机制与还原糖含量变化机制可能存在着部分交叉。     

基于叶片质外体相关成分和酶活性响应的龙葵耐Cd机理研究

为探讨龙葵(Solanum nigrum L.)耐镉(Cd)机理,采用基质培养法,研究了50 μmol·L^-1 Cd(CdCl₂·2.5H₂O)处理120 d对其生长和叶片细胞壁果胶、木质素含量、多聚半乳糖醛酸酶(PG)、纤维素酶(Cx)、NADH-过氧化物酶(NADH-POD)、愈创木酚过氧化物酶(GPOD)和松柏醇过氧化物酶(CAPX)活性以及质外体汁液中过氧化氢(H₂O₂)含量和GPOD、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)和超氧化物歧化酶(SOD)活性的影响。结果表明,与对照相比,Cd处理显著抑制了龙葵地上和地下部的伸长生长、降低了根的干物质积累(P<0.05),但茎的分枝增多,叶面积、叶绿素含量、净光合速率和胞间CO₂浓度无显著变化(P>0.05);细胞壁结合酶NADH-POD、GPOD和CAPX活性以及果胶含量显著增高(P<0.05),而PG活性显著下降(P<0.05),Cx活性和木质素含量无显著变化(P>0.05);质外体汁液中H₂O₂含量显著增加(P<0.05),GPOD和SOD同工酶谱及活性无显著变化(P>0.05);CAT同工酶谱带增宽,APX同工酶谱带数增加,二者活性均显著升高(P<0.05)。说明50 μmol·L^-1 Cd较长时间处理龙葵植株,其地上部分生长仍处于良好状态,表现出对Cd毒害的耐性,这与Cd处理后叶片细胞壁果胶含量和结合酶GPOD以及质外体汁液中APX、CAT活性升高有关,推测这是龙葵耐Cd的一种机理。     

Feeding Solanum glaucophyllum to preparturient multiparous cows prevents postparturient hypocalcemia

Solanum glaucophyllum (SG) contains 1,25‐dihydroxyvitamin D₃ (1,25‐(OH)₂D₃) glycosides. We investigated the effect of SG on hypocalcemia in cows. Serum levels of 1,25‐(OH)₂D₃, total calcium and phosphorus dose‐relatedly increased after feeding with SG, while serum magnesium and chloride levels fell (P < 0.05). We also performed an ethylenediaminetetraacetic acid (EDTA) infusion to induce artificial hypocalcemia. Cows that had been fed 4.0 mg/kg body weight of SG daily for 2 weeks had a higher serum concentration of total calcium at the end of EDTA infusion than those not fed SG (P < 0.05). In a field trial, multiparous cows were assigned to one of four groups: (1) no SG, (2) 1.3 g or (3) 2.6 g of SG daily from 14 days before the estimated calving day until 3 days after calving, or (4) a single feed of 35.75 g SG at 3 days before the estimated calving day. The concentrations of serum total calcium after the calving in each treatment group were (1) 7.4, (2) 7.9, (3) 8.0 and (4) 8.9 mg/dL and higher for (4) than for (1) (P < 0.05). The data suggests that feeding a high dose of SG before the calving may maintain higher concentrations of serum calcium after the calving.