甘蔗原生质体RNA提取条件和方法的优化

为了获得高产量和高纯度的甘蔗原生质体RNA,本研究以新台糖22号（ROC22）和桂糖28号（GT28）甘蔗原生质体为材料,探究了渗透压和原生质体活力对甘蔗原生质体RNA提取效果的影响,并比较了Trizol法、改良Trizol法、改良CTAB法、试剂盒法4种RNA提取方法。结果表明：原生质体渗透压与RNase活性呈显著正相关,原生质体活力与RNase活性呈极显著负相关;RNase活性越高,RNA提取效果越差;使用0.5 mol/L的甘露醇作为渗透压调节剂可获取活力最高的甘蔗原生质体;原生质体活力为70%和90%时所提的RNA完整性好且纯度高,符合后续分子实验的需求,原生质体活力为90%时,RNA产量最高。因此,需要提取甘蔗原生质RNA时,至少需要保证原生质体的活力在70%以上,且原生质体活力越高,RNA的产量越高;采用改良Trizol法,可以大量制备能满足后续分子实验的完整性好且纯度高的RNA,利用该条件和方法,可以确保获得大量高质量的甘蔗原生质体RNA。本研究结果为甘蔗体细胞融合育种过程中的分子生物学研究提供了方法和技术支撑。     

滇东南濒危植物长梗杜鹃的花粉形态及其特性研究

[目的]对滇东南特有濒危植物长梗杜鹃花粉形态及其特性进行系统研究,为长梗杜鹃进一步的遗传育种、花粉种质保存和传粉生物学等相关保护利用提供一些科学依据,并为系统分类研究作旁证。[方法]利用光学显微镜和扫描电子显微镜对长梗杜鹃花朵不同发育时期(花蕾膨大期、花瓣初开期和花瓣盛开期)的花粉形态进行了观察;通过TTC染色法对开花当天不同时段及开花后不同天数的花粉活力进行了快速检测;做L25(53)正交试验探讨10 g·L-1琼脂和不同浓度的蔗糖、H3BO3、Ca Cl2组成的培养基配方对长梗杜鹃花粉萌发的影响。同时,研究了其花粉在不同贮藏温度下活力随贮藏时间的变化情况。[结果]表明:从花蕾膨大期到花瓣盛开期,花粉逐渐成熟,为四合体花粉,呈正四面体排列,四合体直径为43.0 65.4μm,平均51.3μm;单粒花粉近球形且表面有粘丝,具三孔沟。在扫描电镜下,花粉粒外壁为均匀的颗粒状,孔沟周围颗粒细密呈波状突起;在光镜下,花粉粒外壁为内外2层,厚度约相等为0.8 1.2μm。长梗杜鹃整个单花花期花粉活力较强,于开花当天9:00达最高,为92.18%,至花末期(花后第9天)仍保持着48.5%的活力,适宜做父本开展相关杂交育种工作。长梗杜鹃开花当天9:00的花粉在10 g·L-1琼脂+100 g·L-1蔗糖+200 mg·L-1H3BO3+0 mg·L-1Ca Cl2的培养基中萌发率最高,达90.26%;极差分析表明,影响长梗杜鹃花粉萌发率高低的主次顺序为蔗糖→Ca Cl2→H3BO3,一定浓度的蔗糖和H3BO3均起到了很好的促萌效果,而Ca Cl2的添加则显著抑制了其花粉萌发。此外,适宜的低温有利于长梗杜鹃花粉的贮藏,-18℃条件下贮藏48 d仍有一定的萌发力。[结论]长梗杜鹃特殊的花粉外壁纹饰支持了前人对其系统位置划分的合理性。其较高的花粉活力、适宜的固体萌发培养基及贮藏温度的研究为今后以长梗杜鹃为父本的杂交育种提供了可靠的理论依据,对其他同属植物花粉形态及特性研究具有重要的指导意义。     

Investigations on the effect of formalin and iodophor on embryo and larvae development in pikeperch,Sander lucioperca

ABSTRACT

An experiment was conducted to evaluate the effects of incubating pikeperch, Sander lucioperca, eggs in formalin and iodophor solutions for 15 min on embryo survival, the hatching rate, as well as on the rate of misshaped larvae, in order to develop methods for egg surface disinfection. Embryos in the morula stage, in the epiboly stage, and at the beginning of heart beat and blood circulation tolerated formalin concentrations up to 1,500 ppm for 15 min. However, they were very susceptible to iodophor treatment, as >0.1% iodophor solution (=13 ppm active iodine) significantly decreased the percentage of ready-to-hatch embryos and the percentage of hatched larvae. These data of this study recommend the use of formalin at a concentration of up to 1,500 ppm to disinfect pikeperch eggs.     

淡豆豉炮制工艺中加入人参对大豆异黄酮含量的影响

通过比较在传统的淡豆豉(黑豆、黄豆)发酵工艺中加入人参后大豆异黄酮的含量变化,考察人参在淡豆豉发酵过程中对大豆异黄酮含量的影响。以大豆苷、染料木苷和染料木素为测定指标,采用高效液相色谱法进行含量测定。色谱柱为Agilent C18(150 mm×4.6 mm,5μm);流动相为甲醇-水-冰醋酸(40∶60∶0.5);流速0.5 mL.min-1;检测波长260 nm。结果以黄豆为原料发酵的淡豆豉,加入人参后大豆苷和染料木素的含量显著升高,染料木苷显著降低;以黑豆为原料发酵的淡豆豉,加入人参后大豆苷的含量显著降低,染料木苷和染料木素的含量显著升高。因此,淡豆豉传统发酵工艺中加入人参后,大豆异黄酮含量变化因原料不同而异。     

Relative importance of chinook salmon abundance on resident killer whale population growth and viability

相似文献   

Effects of variable egg incubation temperatures on the embryonic development in Arctic charr Salvelinus alpinus

An evaluation of different thermal regimes during incubation of Arctic charr Salvelinus alpinus L. egg was conducted with regard to embryo survival and development. The study consisted of two major parts: In experiment A, a range of commonly used thermal regimes, constant and variable ranging from 2.7°C to 7°C, were tested for differences in embryo survival and developmental status of newly hatched alevins. Experiment B assessed the timing of a shift from autumn (6°C) to winter temperature (2.5°C) and its effect on embryo survival and the occurrence of spinal deformities in newly hatched alevins. In general, incubations that were initiated at temperatures below 3°C resulted in significantly higher mortality rates and spinal deformity frequencies than other treatments. Hence, Arctic charr eggs require an initial warmer period for unimpaired embryonic development. A general negative correlation was found between the duration of this warmer period (6°C) and both mortality rates and frequency of spinal deformities. The total absence of the 6°C period had the most detrimental effect, with deformity frequencies at least four times higher than all treatments with an initial warmer period. The upper incubation temperature threshold was not reached in the study. The highest temperature tested (7°C) did not result in excess mortality or increased deformity frequency. Higher incubation temperature resulted in less developed alevins at hatching, i.e. alevins were smaller with more yolk remaining. However, measurements on alevins from incubations at variable temperatures revealed different temperature effects on growth and yolk consumption during different parts of embryonic development.     

Modelling the population viability of a threatened amphibian with a fast life‐history

相似文献   

6个蓝莓品种花粉特性和花粉育性的比较

为了探究湖南蓝莓引种区不同品种的花器形体特征及花粉生活力,从而为南方地区蓝莓引种栽培和授粉树的配置提供参考依据,采用纤维素酶溶解法、联苯胺染色法和离体培养等技术,对‘莱格西’、‘布里吉塔’、‘夏普蓝’、‘密斯梯’、‘粉蓝’、‘梯芙蓝’等6个蓝莓品种的花粉特性、花粉量、花粉生活力、花粉萌发率等进行了比较研究。测定结果表明:1蓝莓花粉量在各品种间存在差异,‘莱格西’、‘布里吉塔’、‘夏普蓝’、‘密斯梯’、‘粉蓝’、‘梯芙蓝’的花粉数依次为162、250、178、166、141、170粒/花药,其中‘布里吉塔’的花粉数量最多,‘夏普蓝’次之,‘粉蓝’最少;2蓝莓花粉生活力在各品种中存在差异,‘莱格西’、‘布里吉塔’、‘夏普蓝’、‘密斯梯’、‘粉蓝’、‘梯芙蓝’的花粉生活力依次为30.87%、26.79%、31.34%、23.83%,18.80%、24.83%,其中‘夏普蓝’的花粉活力最大,‘莱格西’次之,‘粉蓝’最小;3 6个蓝莓品种花粉萌发所需的最适蔗糖浓度不尽相同,‘莱格西’、‘布里吉塔’、‘密斯梯’在蔗糖浓度为10%时其花粉萌发率均最高,依次为25.60%、9.10%、27.20%;当蔗糖浓度为15%时,‘夏普蓝’、‘粉蓝’、‘梯芙蓝’3个蓝莓品种的萌发率均达到最高值,依次为26.20%、21.20%、61.40%;4 6个蓝莓品种中,‘夏普蓝’花粉萌发所需的最适硼酸浓度为300μg·L-1,而其他5个品种花粉萌发的最佳硼酸浓度均为200μg·L-1;5 6个蓝莓品种花粉萌发所需的最适培养温度均为25℃。研究结果表明:6个蓝莓品种中,‘莱格西’和‘夏普蓝’均较适合用作授粉植株,其花粉量和花粉萌发率均较高。     

Retinoids,retinoid analogs,and lactoferrin interact and differentially affect cell viability of 2 bovine mammary cell types in vitro

Two bovine mammary cell types (BME-UV1 and MeBo cells) were used to evaluate the effect of natural retinoids, retinoid analogs, and bovine lactoferrin (bLf) on cell viability in vitro. Experiments with Alamar Blue showed a linear relationship between fluorescence and cell viability index. The BME-UV1 cells exhibited twice the metabolic activity but required half the doubling time of the MeBo cells. The BME-UV1 cells were very sensitive to all-trans retinoic acid (atRA) inhibition of cell viability (P < 0.05) and exhibited a dose-dependent inhibition with 9-cisRA (9cRA; P < 0.05). The MeBo cells exhibited some inhibition with these natural ligands (P < 0.05), but they were not as sensitive. The addition of bLf had similar inhibitory effects (P < 0.05) on cell viability of the 2 mammary cell types. Applications of RA receptor (RAR) agonist indicated that the stimulation of the RAR in both mammary cell types was highly effective in inhibition of cell viability (P < 0.05), whereas the application of an RAR antagonist stimulated MeBo cell viability (P < 0.05) and inhibited BME-UV1 cell viability (P < 0.05). Finally, the use of the RAR antagonist in conjunction with bLf indicated a rescue of the bLf effect in the MeBo cells, suggesting that bLf is acting through the RAR receptor. Conversely, bLf reverted inhibition of cell viability by 9cRA in the BME-UV1 cell type (P < 0.05). We conclude that RAR interaction in bovine mammary cell types regulates cell viability in vitro; we hypothesize that the natural ligands mediate regulation of bovine mammary cell viability in vivo and that bLf can either enhance or reverse the retinoid-induced inhibition of cell viability, depending on the type of bovine mammary cell studied.     

凉州区肉牛繁殖技术试验示范与推广研究

[目的]为筛选出适合凉州区肉牛繁殖的新模式。[方法]选择夏洛莱（XL）、利木辛（LM）、安格斯（AG）成年种公牛生产冻精,在西门塔尔和本地黄牛杂交一代（西本F1）成年母牛上实施人工授精,对杂交犊牛分月龄进行外貌体尺测定。[结果]XL冻精与西本F1代的后代体重、体高、体斜长、胸围、管围等生长指标均高于LM和AG后代指标。试验2方法是选择XL、LM、AG成年母牛生产冻胚,在成年母牛上实施胚胎移植,犊牛出生后进行外貌体尺测定。结果是：夏洛莱胚胎移植后代牛初生重、6月龄、12月龄体重、体高、体斜长、胸围、管围均高于利木辛与安格斯牛杂交后代（P〈0.01）。试验3方法是选择西杂F1和F2代母牛,用引进的XM（西门塔尔）牛X性控冻精实施人工授精,犊牛出生后统计相关数据。结果是：三组共授配母牛138头,情期妊娠75头,情期平均受胎率为54.1%,平均母犊率为90.3%。[结论]夏洛莱品种改良西杂一代牛后代生长发育比其余品种好;实施胚胎移植后,夏洛莱品种胚胎移植后代牛比其余品种在凉州区生长适应性强,适宜在该环境条件下生长;西门塔尔牛性控冻精的配种试验是成功的,是生产母犊的较好方法。