Studies on In Vitro Flowering and Fruiting of Perilla frutescens

Influences of PGR, sucrose, and ammonium nitrate on in vitro flowering and fruiting from cotyledon explants of P. frutescens were studied. The regenerated shoots at 2-4. cm from cotyledon explants on MS medium supplemented with 0.5 mg L^-1 BA and 1.0 mg L^-1IAA were excised and transferred to MS medium containing 30 g L^-1 sucrose, 8.25 g L^-1 ammonium nitrate, and 1.0 mg L^-1 BA. Following 40 d of culture, 86.2% of them flowered and set seeds. These seeds were germinable and developed into flowering plants in the fields. This study provides a simple system for rapid breeding of P. frutescens and studying the physiological mechanism of flowering of plants.     

应用体外法评定科威418的营养价值

科威４１８是作者通过特殊工艺研制而成的牛羊高蛋白复合添加剂，其中含有尿素、营养保护剂、酶及多种微量元素等。体外试验表明，以豆饼、尿素及科威４１８作为氮素饲料时，培养３ｈ的培养液中ＮＨ＿３Ｎ浓度依次为１４．５，２８．７及２２．３ｍｇ·（１００ｍＬ）￣（－１）（Ｐ＜０．０１）；培养１２ｈ的ＮＨ＿３Ｎ浓度依次为７．３，１１．４及１３．３ｍｇ·（１００ｍＬ）￣（－１）（Ｐ＜０．０５）；真蛋白氮浓度依次为３８．６，２９．８及３５．４ｍｇ·（１００ｍＬ）￣（－１）（Ｐ＜０．０５）；基质风干物消化率依次为５４．９７％，６１．４４％及６４．９１％（Ｐ＜０．０１）。     

牛磺酸对昆明小鼠早期胚胎体外发育和着床的影响

本研究主要探讨了牛磺酸对昆明(KM)小鼠体外胚胎发育及对胚胎着床的作用。在mKSOM(modified potassium simplex optimized medium)胚胎体外培养液中分别添加不同浓度牛磺酸,培养24、72 h后分别统计4-细胞率、囊胚率、囊胚总细胞数;采用子宫角注射的方法,研究牛磺酸转运抑制剂β-丙氨酸对孕D9胚胎着床数的影响。结果表明,添加10 mmol/L牛磺酸培养液组的胚胎4-细胞率高于对照组(P<0.05),囊胚率与囊胚总细胞数明显高于对照组(P<0.01)。子宫角注射β-丙氨酸(20 μg/mL)显著降低D9着床的胚胎数。本研究初步明确了牛磺酸对小鼠体外胚胎发育及对胚胎着床的影响,为进一步研究胚胎发育和着床提供参考。     

小鼠心肌细胞的体外培养

对采用组织块法和酶消化法体外培养小鼠心肌细胞的结果进行了比较,并对体外培养心肌细胞的生理活性进行了初步测定。结果表明,用组织块法分离培养胎、乳鼠心肌可得到自主节律性搏动的心肌细胞;分别用1g/L胶原酶+10g/LBSA,0.5g/L胰酶+1g/L胶原酶,1g/L胰酶分离培养胎、乳鼠心肌,均可得到大量心肌细胞,且细胞活力较好,能自主博动20d以上;用上述2种方法分离培养成年鼠心肌,结果均不理想;在载有自主节律性搏动的小鼠心肌功能性细胞团的培养皿中加入Ca2+和肾上腺素后,细胞团搏动速度加快,搏动力加强;加入K+后则细胞团搏动速度减缓,搏动力减弱,其对钾、钙、肾上腺素的反应与在体内基本一致,表明体外培养的小鼠心肌细胞具有与在体心肌细胞相似的功能。     

木棉纤维的基本结构和性能

目的进一步完善木棉的组培快繁体系,实现木棉组培苗不定根的定向诱导,提高木棉生根苗的移栽成活率。方法以木棉组培丛生芽为材料,通过激素、纵向划痕和暗培养等3种处理促进根系诱导,比较不同处理下木棉组培苗的生根率、根系数量和移栽成活率,同时对生根过程进行显微观察。结果在单独添加NAA和IBA时,最高生根率分别为19.66%和39.15%;而NAA与 IBA组合处理的生根率可达95.80%,优于两者单独添加处理,且生根苗以1条不定根为主。纵向划痕处理以2~3个划痕效果较好,其生根率可达94%以上,生根苗以2条不定根为主。暗培养处理以6 d最佳,生根率为97.36%,生根苗以1条不定根为主。显微观察发现：木棉组培苗生根属于皮层生根类型。对1、2和3条不定根等3种类型的生根苗进行移栽,其移栽成活率之间无显著差异(P>0.05),但生物量存在显著差异(P<0.05),其中3条不定根的移栽苗生物量最大。结论本研究优化了木棉组培苗的生根条件,为木棉组培苗的移栽炼苗以及不定根的定向调控提供了理论参考。     

马铃薯耐盐突变体的离体筛选

以4个马铃薯栽培品种的叶片为外植体进行耐盐突变体筛选,结果表明:不同品种愈伤组织的耐盐性存在差异,东农303、鄂1号两品种在细胞水平对NaCl的最大耐受质量浓度为8g/L,费乌瑞它为6g/L,夏波帝为4g/L。当继代次数低于10次,叶片耐盐愈伤组织在无盐胁迫诱导分化培养基上才有进一步诱导再生植株发生的能力;而在盐胁迫诱导分化培养基中,愈伤组织继代次数超过8次即很难继续分化再生。     

Effect of PMSG/hCG Superovulation on Mouse Embryonic Development

Kunming mouse strain is widely used in China, and the superovulation was administrated with 10 IU PMSG combined with 10 IU hCG. In this study, the effects of the exogenous gonadotropins on superovulation of Kunming mice and embryo quality derived from the superovulated mice were assessed. Female mice at 6-8-wk old were superovulated with 0, 5, 7.5 and 10 IU PMSG/hCG and mated with male mice. The embryos were retrieved at 2.5 d post coitum. No statistic difference was observed for the number of 2-cell embryos collected per mouse between control and 5 IU PMSG/hCG treatment group, but the number significantly increased for 7.5 and 10 IU PMSG/hCG treatment group (P<0.05). The average number of 4- cell and 8-cell embryos collected from each mouse significantly differed between control and 5, 7.5, 10 IU PMSG/hCG treatment groups (P<0.05). When 8-cell embryos derived from mice administrated with 0, 5, 7.5 and 10 IU PMSG/hCG were cultured in KSOM, the blastocyst development rates were 88.1, 94.7, 96.1 and 94.3%, respectively, which were similar to control (P>0.05). This indicated that exogenous gonadotropins have no effects on development of Kunming mouse embryos. The quality of blastocyst was assessed by labelling with Hoechst and propidium iodide for inner cell mass and trophectoderm cells, the result showed that ICM/TE ratio significantly decreased for 10 IU PMSG/hCG treatment group compared with control, 5 and 7.5 IU PMSG/hCG treatment group (P<0.05). This suggested that the embryo quality of Kunming mouse has been affected by high dose of gonadotropins.     

不同培养条件下小鼠体外胚泡着床模型的研究

用对比的方法观察不同培养条件对小鼠体外胚泡着床的影响,从而建立在体外研究着床机制的简易着床模型。在无血清单独培养系统、子宫内膜上皮细胞(EEC) 无血清共培养系统、EEC 3%胎牛血清(FCS)系统和EEC 0.4%牛血清白蛋白(BSA)系统中分别移入胚泡,观察并统计脱带率、粘附率和扩展率。结果显示后面两组的脱带率、粘附率、扩展率明显高于前面两组,差异显著(P<0.05);而EEC 3?S共培养组的脱带率、粘附率、扩展率和EEC 0.4%BSA共培养组相比,差异不显著(P>0.05)。因此胚泡与子宫内膜上皮细胞用FCS共培养与用BSA共培养都是比较理想的着床模型,可以用来体外研究着床的影响因素。     

胡椒木叶片离体培养与植株再生

探讨了胡椒木(Zanthoxylum piperitum)叶片的离体培养.选取胡椒木幼嫩叶片作为外植体,接种在不同激素组合的培养基上进行诱导培养,结果表明,最适的诱导培养基为MS+6-BA1.0 mg/L+NAA0.7 mg/L,分化率为91.7%,最适的增殖继代培养基为MS+6-BA1.0 mg/L+NAA0.7 mg/L,所产生的不定芽在1/2MS+ NAA0.4 mg/L的培养基上可全部生根,长成完整植株.首次证明以胡椒木的叶片为外植体可以进行快速离体培养.     

茎直黄芪提取物的体外抑菌作用

为研究茎直黄芪提取物的体外抑菌作用,采用琼脂扩散和试管稀释法对其抑菌效果进行了研究。结果显示茎直黄芪石油醚部分、氯仿部分、乙酸乙酯部分和正丁醇部分提取物对金黄色葡萄球菌、大肠埃希菌、无乳链球菌、沙门氏菌和多杀性巴氏杆菌等5种细菌有不同程度的抑菌作用,其中正丁醇部分抑菌作用较强。通过进一步试验表明正丁醇部分主要抑菌活性成分集中在20%乙醇和40%乙醇部分,尤其以后者抑菌活性最佳。40%乙醇部分对5种细菌的MIC均为6.25 mg/mL,对大肠埃希菌的MBC为6.25 mg/mL。对金黄色葡萄球菌、沙门氏菌和多杀性巴氏杆菌的MBC均为12.25 mg/mL,对无乳链球菌的MBC为25.00mg/mL。通过研究发现茎直黄芪提取物在体外具有明显的抑菌作用。     

Plant Regeneration from In Vitro Cultured Hypocotyl Explants of Euonymus japonicus Cu zhi

Adventitious shoots were successfully regenerated from hypocotyl explants of in vitro cultures of Euonymus japonicus Cu zhi. Hypocotyl slices were cultured on Murashige and Skoog （MS） and B5 basal medium supplemented with varied concentration of different plant growth-regulators, e.g., α-naphthaleneacetic acid （NAA） and indole-3-butyric acid （IBA） in combination with 6-benzylaminopurine （6-BA） and kinetin. The study showed that shoots could be directly regenerated from hypocotyl explants without the intervening callus phase; MS medium was more suitable for adventitious shoots regeneration. The ability of hypocotyls segments to produce shoots varied depending upon their position on the seedlings. The highest regeneration rate was obtained with hypocotyl segments near to the cotyledon cultured on MS basal medium supplemented with 1.5 mg L^-1 6-BA and 0.05 mg L^-1 NAA （63.64%）. The regenerated shoots were readily elongated on the same medium as used for multiplication and rooted on half-strength MS basal medium supplemented with 1.0 mg L^-1 IBA and 100 mg L^-1 activated carbon. After being transferred to greenhouse conditions, 96% of the plantlets were successfully acclimatized. This regeneration system is applied for genetic transformation now.     

朝鲜淫羊藿种胚组培研究

【目的】建立朝鲜淫羊藿（Epimedium koreanum Nakai）种胚组织培养快繁体系,筛选其适宜的组织培养条件。【方法】以朝鲜淫羊藿种胚为外植体,研究外植体最佳消毒方式;在1/2MS基础培养基上,采用L₉(3⁴)正交试验设计,探讨6-BA、NAA、IBA和蔗糖对种胚愈伤组织诱导的影响,筛选适宜培养基,并分析不同激素组合对愈伤组织增殖、丛生芽增殖及幼芽生根的影响。【结果】朝鲜淫羊藿的最佳消毒方式为：利用体积分数0.1% HgCl₂对种子表面消毒8 min,再对种胚消毒5 s,该条件下种子存活率为71.11%;诱导种胚愈伤组织的适宜培养基为1/2MS+1.0 mg/L 6-BA+0.5 mg/L NAA+0.5 mg/L IBA+30 g/L蔗糖,诱导率为47.78%;愈伤组织增殖的适宜培养基为1/2MS+0.5 mg/L KT+0.3 mg/L NAA,增长率为172.67%;丛生芽的适宜增殖培养基为1/2MS+2.0 mg/L 6-BA+0.4 mg/L NAA,增殖系数为2.78;0.5 mg/L NAA的生根效果最好,生根率为43.33%。【结论】建立了朝鲜淫羊藿种胚适宜的组织培养体系,在该体系下,组培苗长势良好,但生根效果需进一步研究。     

鸡胚原壳体外培养研究

以种蛋原壳为鸡胚体外培养的孵化蛋壳,就开孔位置、开孔大小以及鸡胚胚龄对鸡胚体外培养效果的影响进行了研究。结果表明:(1)小头侧面开孔明显优于小头顶端开孔(P<0.01),小头侧面开孔的已孵化3 d鸡胚的孵化率为80.0%,而小头顶端开孔的则只有60.0%;(2)随着开孔直径的增加,鸡胚孵化率呈下降趋势,开孔直径为0.5,1.0和1.5 cm时的孵化率分别为80.0%,30.0%和16.7%;(3)在小头侧面开孔且孔直径为0.5cm时,X期鸡胚和已孵化3 d鸡胚的体外培养孵化率差异达极显著水平(P<0.01),已孵化3 d鸡胚的体外培养孵化率为80.0%,X期鸡胚的孵化率则为0。     

Expression of Eph-Ephrin A Molecules in Endometrium During Swine Embryo Implantation Examined Using Real-Time RT-PCR

Erythropoietin-producing hepatocellular receptor and its membrane-bound ligands (Eph-Ephrin) system could regulate some mammalian blastocyst attachment and spreading. In order to investigate the involvement of the Eph-Ephrin system in swine embryo attachment, mRNA expression of Eph-Ephrin molecules in endometrium was examined by real-time RT-PCR during embryo implantation in pigs. The results indicated that mRNA expressions of Eph A5, A7 and Ephrin A5 all continually increased from pregnancy day 13 to 24. Ephrin A3 mRNA expression significantly increased from day 13 to 18 and decreased from day 18 to 24, and the expression was the lowest on pregnancy day 13 and the highest on day 18. However, Ephrin A4 mRNA expression was the lowest on pregnancy day 18 and the highest on day 24, and the expression decreased from day 13 to 18 and increased from day 18 to 24. Furthermore, mRNA expressions of Eph A5 and A7 were both found in other tissues, such as brain, muscle, intestine, stomach, etc. These findings suggest that the Eph-Ephrin system may play an important role in regulating the contact between blastocysts and endometrium during swine embryo implantation.     

HA对牛卵母细胞体外成熟和早期胚胎体外发育影响的研究

在TCM-199中添加一些确定的辅助成分,探讨在无血清培养基中添加不同浓度HA对牛卵母细胞体外成熟及早期胚胎体外发育的影响。结果表明,在卵母细胞成熟培养液(TCM-199-m)中添加4.0mg/mL的HA时,卵母细胞的成熟率和卵裂率与对照组(BSA)无显著差异(P>0.05),分别为85.14%,83.57%和89.47%,85.24%,但显著高于其他各浓度组(P<0.05),表明HA在卵母细胞无血清成熟培养中可代替BSA。在受精卵体外培养液(TCM-199-c)中添加HA时,囊胚发育率与对照组(BSA)差异不显著(P>0.05),分别为27.64%和26.51%,但显著高于TCM-199-c组(P<0.05),表明HA对牛胚胎体外发育具有明显的促进作用,在无血清培养中可以代替BSA。当在TCM-199-c+HA中添加少量的BSA时,囊胚发育率(31.19%)显著高于其他各组(P<0.05),表明HA欲完全代替BSA还有不完善之处,有待进一步研究。     

花椒麻素在大鼠肠道的吸收动力学研究

【目的】研究花椒麻素在大鼠肠道的吸收动力学特征。【方法】以SD大鼠为供试动物,建立大鼠体循环肠灌流模型,并采用建立的高效液相色谱法(HPLC)测定肠循环液中花椒麻素质量浓度,研究花椒麻素在大鼠肠道不同部位(十二指肠、空肠、回肠、结肠)的吸收特性,分析不同质量浓度花椒麻素对大鼠肠吸收速率常数(Ka)、吸收百分率(P)和吸收半衰期(t1/2)的影响。【结果】25~100μg/mL花椒麻素在大鼠肠道的Ka为30.0~65.0min-1,P为30.16%~56.81%,t1/2为100.10~232.09min。花椒麻素在各肠段的P和Ka均随其质量浓度的增加而增大,而t1/2随其质量浓度的增加而减少。花椒麻素Ka和P均按空肠、回肠、十二指肠、结肠顺序依次下降,而t1/2依次升高。【结论】花椒麻素在大鼠肠道的吸收过程属于一级动力学过程,属被动转运,吸收部位主要是空肠和回肠。     

云南松胚性愈伤组织诱导研究

以未成熟云南松种子为研究材料,选择合适的消毒方法,进行合子胚发育状态观测,从外植体类型、采果母树基因型、培养基种类3方面探讨其对胚性愈伤组织诱导的影响,并鉴定了胚性愈伤组织。结果表明：75%乙醇消毒30 s,无菌水冲洗2次,2%氯酸钠浸泡15 min,无菌水冲洗4～5次为种子最佳消毒方案;不同外植体类型对出愈率影响不显著,但对胚性愈伤组织的形成具有极显著影响,取带胚乳的合子胚作为外植体的胚性愈伤组织诱导率可达34.3%,而合子胚作为外植体时诱导率为14.3%;采用14种培养基接种,平均诱导率在2.2%～17.6%之间,其中接种于MLV+2, 4-D 2.0 mg/L+6-BA 1.0 mg/L+KT 1.0 mg/L+肌醇 0.1 g/L,1/2LM+2, 4-D 2.2 mg/L+6-BA 1.1 mg/L +AgNO₃ 3.4 mg/L+肌醇 0.1 g/L,DCR+6-BA 0.63 mg/L +KT 0.61 mg/L+NAA 2.0 mg/L+肌醇 0.2 g/L+麦芽糖15 g/L培养基的胚性愈伤诱导率分别为17.6%、17.6%、16.5%,表明该3种培养基适合于云南松的胚性愈伤诱导;8株云南松优株未成熟带胚乳合子胚的胚性愈伤组织平均诱导率为5.1%～19.4%,最高的是076号云南松,最低的是075号云南松,表明基因型对胚性愈伤组织诱导具有极显著的影响。     

高光强下蔗糖浓度对姜试管苗生长及相关生理特性的影响

以河南张良姜和四川竹根姜常规条件下繁殖的试管苗为试材,研究了120μmol.m-2.s-1光强下不同蔗糖浓度(0%、1%、3%)对其相关生理特性的影响。结果表明,河南张良姜和四川竹根姜试管苗的茎叶鲜重、根鲜重、单株鲜重和增殖系数随蔗糖浓度降低而降低,根与茎叶的可溶性糖和淀粉含量也均呈现下降趋势,根的可溶性糖和淀粉含量比茎叶中下降更为迅速,其生长受到了更大限制,根冠比减少。随蔗糖浓度降低,叶片中叶绿素和类胡萝卜素含量均降低,气孔长度和宽度显著增加,气孔密度变小。     

“三维一体”运训模式的构建与实践研究*

坚持现代体育“以人为本”的教育理念,彰显体育专业的实践特色,走教学与运训相结合的发展之路,做到课内与课外相结合,实训与竞赛相结合,校外与校内实践相结合,实现课堂教学、课外活动、社会实践有机的结合,培养与发展学生的专业技能和实践能力,是当前地方本科体育院系体育改革热的。研究立足于学生主体性的发挥,发展学生的个性、创造性、独立性和协作能力,构建新的运训模式,旨在全面提高体育专业的教学质量和人才培养质量。