代谢互补抑制剂碘乙酰胺和罗丹明对3种豆科牧草原生质体活力和再长的影响

研究了不同浓度代谢互补抑制剂碘乙酰胺(IOA)和罗丹明6G(R-6G)处理与里奥百脉根(Lotuscorniculatus L.cv.Leon)、扁蓿豆(Melilotoides ruthenica(L.)Sojak)及清水紫花苜蓿(Medicago sativa L.cv.Qingshui)3种豆科牧草愈伤组织原生质体活力和再生力之间的相关性。结果表明,3～10mmol/L的IOA和40～70μg/ml的R-6G分别处理10min,可使3种豆科牧草原生质体的活力及植板率明显下降,各处理浓度与原生质体活力和植板率之间均存在极显著的负相关(p<0.01)。培养第7天,所有处理均可见再生的小细胞团;培养30～40d,抑制剂临界浓度下原生质体丧失形成愈伤组织的能力。IOA对原生质活力的抑制作用更明显,R-6G对植板率的影响更显著,并可使原生质体发出红色荧光,有利于细胞融合时亲本原生质体的标识。适宜3种豆科牧草的抑制剂及临界浓度分别是:百脉根5mmol/L IOA或50μg/ml R-6G;扁蓿豆7mmol/L IOA或70μg/ml R-6G;清水紫花苜蓿3mmol/L IOA或40μg/mL R-6G。     

Characteristics and Warning Indexes of Rice Seeds Viability Loss During Storage at 45℃ Constant Temperature

Seed aging characteristics of rice was investigated in this study. Seeds of 34 japonica rice (O-ryza sativa subsp. japonica) varieties were held at 45℃ constant temperature. Changes in seed viability and seed vigor during aging process were measured to study seed viability-losing characteristic and to determine warning index for seed viability loss. As a result, seed viability survival curves were obtained across different rice accessions at 45℃ constant temperature. The curves appeared to be contra-sigmoid survival curves. The loss of seed viability in the aging process consisted of two phases. The first phase took a long duration, in which the viability of vigorous seeds declined slowly. In the second phase, seed viability declined rapidly. It was obvious that seed viability declined inconsistently during storage. It also showed that seed germination was prolonged and the seedling was significantly weakened before the coming of the rapid declining phase of seed viability. These two parameters could be used to indicate seed quality during storage. The rate of compatibility of tests (RCT), coefficient of variation (CV), vigor of seedling, the day the seeds start to germinate could be used as warning indexes to indicate overall quality of a mass of accessions. These warning indexes could also be used in monitoring the viability of seeds stored in the seed genebank.     

比较MTS与MTT用于大鼠热应激模型中肠上皮细胞活力检测

[目的]MTT法与MTS法线性关系比较与热应激模型筛选.[方法]以IEC-6及SF9细胞为研究对象,取对数生长期细胞,设置4个接种浓度,将两种细胞制成对应浓度细胞悬液接种于96孔板24 h,分别加入两种试剂后在490 nm处读取OD值.根据OD值确定两种细胞最佳接种浓度.并根据OD值比较两种检测细胞活力方法哪种更加适合于动物贴壁细胞IEC-6细胞活力测定.取最适浓度的IEC-6细胞加入MTS后孵育1.5、2.5、3.5h三个不同时间点检测OD值,根据OD值确定最佳孵育时间.将IEC-6细胞在最佳接种浓度下接种于96孔板24 h,分别在43、41、39℃下采用不同热应激时间4、8、12、24 h,测定OD值,根据OD值确定IEC-6细胞热应激模型.[结果]MTS法在动物贴壁细胞IEC-6及植物半贴壁细胞SF9中检测的准确度度高,灵敏度好,是比MTT法更优的检测贴壁细胞增殖活性的方法.IEC-6细胞最佳接种浓度为4.1×104/mL至5.5×104/mL,SF9细胞最佳接种浓度为3.28×105/mL至4.4×105/mL.MTS最佳孵育时间为3.5h,并确定大鼠空肠上皮细胞热应激模型为43℃,6h.[结论]检测贴壁不牢的细胞活力时MTS法优于MTT法,并成功建立IEC-6细胞热应激模型.     

F-2毒素对体外培养猪卵巢颗粒细胞的毒害及V-E的解毒效果

采用细胞体外培养的方法,在卵巢颗粒细胞对数生长期,对F-2毒素的繁殖遗传毒性和V-E对其的解毒作用进行了研究．结果表明,各毒素组细胞活性显著低于正常对照组,而丙二醛（MDA）含量显著低于正常对照组;各解毒组细胞活性均显著高于各毒素组,但明显低于正常对照组,MDA含量显著低于各毒素组,但明显高于正常对照组．这表明,F-2毒素可能是通过脂质过氧化作用,对颗粒细胞致毒,V-E对F-2毒素具有较好的解毒作用,能提高中毒卵巢颗粒细胞的抗过氧化能力．     

复方中草药制剂对新西兰兔精液品质的影响

选取6～7月龄新西兰公兔12只,随机分为试验组和对照组,每组6只,研究以淫羊藿、菟丝子、熟地黄等为主要成分的复方中草药制剂对新西兰公兔精液品质的影响。结果表明:实验组公兔在饲喂该复方中草药制剂8～14和15～21d内的射精量、精子密度和活率等指标与对照组公兔精液品质相比差异极显著(P<0.01);停药后10d内,实验组和对照组公兔的射精量差异显著(P<0.05),精子密度和活率差异极显著(P<0.01);但是整个试验过程中精子畸形率两组间差异不显著。说明该复方中草药制剂可以用来改善公兔的精液品质,有利于生产实践中进一步发挥优良公兔的种用价值。     

百日草柱头可授性和花粉生活力的研究

 【目的】观察百日草花后柱头的形态变化，测试百日草的柱头和花粉的寿命，为提高百日草的人工授粉效率和杂交育种提供依据。【方法】采用联苯胺－过氧化氢测定柱头可授性、花粉离体萌发试验和田间试验相结合的方法进行研究。【结果】（1）百日草在8:30～16:00开花，花粉生活力为18 h；（2）舌状花柱头保持膨胀，花柱挺立可达7～10 d；（3）联苯胺－过氧化氢测定柱头可授性的结果显示，花后1～3 d可授率为79.9%～83.1%，4～6 d为56.1%～63.0%，7～10 d为37.0～43.3%；（4）丫状时期、丫状尖端稍弯曲时期、柱状时期田间授粉的结实率分别为76.3%、48.5%和36.8%；（5）未经处理的柱头结实率为48.8%，去丫状一裂者为30.9%，去柱头只留花柱者为19.9%。【结论】（1）用联苯胺－过氧化氢方法测定百日草柱头可授性可行；（2）花后2～3 d柱头丫状时期进行人工授粉结实率高；（3）柱头顶端的毛状突起增大了吸收花粉的表面积，有利于授粉结实。     

不同培养基组分对5个蜡梅品系花粉萌发和花粉管生长的影响

以河南省许昌市3个类型5个品系的蜡梅花粉为试验材料,研究了不同培养基组分对蜡梅花粉萌发和花粉管生长的影响,以筛选出适宜不同类型蜡梅花粉萌发的最佳培养基,为生产实践中科学授粉和杂交育种提供理论依据.结果表明:聚乙二醇是蜡梅花粉萌发和花粉管生长的关键因素,当培养基中没有聚乙二醇时,花粉不能萌发;蔗糖不是蜡梅花粉离体萌发的必...     

响应面法优化酶法提取沙苑子三萜的工艺及抗氧化活性研究

旨在研究酶法辅助技术提取沙苑子三萜的工艺及体外抗氧化活性。在单因素实验的基础上,确定液料比、酶添加量、酶解温度、酶解时间4 个因素的Box-benhnken 的实验设计,以三萜的提取率为响应值,采用响应面法优化沙苑子三萜的提取工艺,建立并分析各因素与指标值的数学模型;采用自由基清除能力体系评价沙苑子三萜的抗氧化活性。优化分析所得的最佳工艺参数为：液料比为40 mL/g,酶添加量为500 μg/mL,酶解温度为40℃,酶解时间为50 min,三萜提取率理论值为8.08%,实际值为7.98%,其RSD 为0.12%。沙苑子三萜对DPPH自由基、O2 -·自由基、羟自由基及Fe3+具有较强的清除作用。因此,该方法高效、简单,可用作沙苑子三萜的提取;沙苑子三萜具有明显的体外抗氧化活性。     

GDC-0032 inhibits cell viability and induces DNA damage in U251 human glioma cells

AIM: To investigate the effect of phosphatidylinostiol 3-kinase (PI3K) inhibitor GDC-0032 on cell viability, cell cycle and DNA damage in human glioma U251 cells. METHODS: The cell viability was analyzed by MTT assay. The cell cycle distribution of U251 cells was examined by flow cytometry. The protein expression was examined by Western blot. The expression and localization of γ-H2AX were determined by laser-scanning confocal microscopy. RESULTS: GDC-0032 inhibited the cell viability in a dose-dependent manner. U251 cells showed G₁ phase arrest accompanied with upregulation of p27 protein after exposure to GDC-0032, while the expression of cell division cycle protein 2 (Cdc2) was inhibited. GDC-0032 treatment increased the formation of γ-H2AX foci and histone H2AX phosphorylation in the U251 cells. In addition, GDC-0032 upregulated the phosphorylation levels of mitogen-activated protein kinases, including extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), in the glioma cells, while the expression of survivin was inhibited. CONCLUSION: GDC-0032 inhibits U251 cell growth by inhibition of cell viability and cell cycle progression. GDC-0032 also induces DNA damage of U251 cells. The anticancer activity of GDC-0032 is associated with increasing the activity of ERK and JNK and downregulating the expression of survivin.     

Gametophytic self-incompatibility causes seedlessness in ‘Kagzi Kalan’ lemon (Citrus limon)

The present study aimed to study the flower and pollen structure, pollen viability, in vitro pollen germination of ‘Kagzi Kalan’ lemon (Citrus limon (L.) Osbeck), and in vivo pollen tube growth in self- and cross pollination to understand the mechanism of seedlessness. The results revealed that Kagzi Kalan had tetra-colporate spherical shaped pollen of uniform size. The results of Acetocarmine and FDA test and in vitro germination indicated normal pollen viability and germinability in Kagzi Kalan. In vivo pollen tube growth showed > 50% pollen germination on stigma in all treatments though a greater number of pollen tubes at the mid style, base of the style, and near the ovule was found in cross pollination. Contrary to this, despite optimum pollen germination (53.51–70.37%) in self-pollination treatments, none of the pollen tubes reached the base of the style due to abnormal growth of pollen tubes, viz. curvatures, inhibition of pollen tube growth, and callose deposition. The inhibition of pollen tubes at the mid style under self-pollination treatments, and production of seeded fruits under cross pollination treatments, indicate the presence of self-incompatibility and the site of inhibition of pollen tubes. Thus, gametophytic self-incompatibility (SI) caused seedlessness in Kagzi Kalan by blocking fertilization in the ovary.