朱顶红无菌苗叶片高效再生体系

以朱顶红(Hippeastrum vittatum)叶片为外植体进行离体培养,具有取材方便、试材充足、成本低等优势,但叶片诱导再生率极低,是朱顶红离体培养的一大难题。本试验中分别以‘花孔雀’和‘黑天鹅’朱顶红无菌苗叶片为外植体,探究了不同植物生长调节剂和不同取材部位对不定芽诱导和继代增殖的影响。结果表明:最佳外植体为MS培养基中培养10 d形成的幼嫩叶片基部(0.5 cm),在光照16h·d~(-1)(光照强度36μmol·m~(-2)·s~(-1))下,不定芽诱导的最适培养基为MS+2 mg·L~(-1) 6-BA+1 mg·L~(-1) NAA+2 mg·L~(-1) TDZ,两个品种的不定芽均以间接途径发生,其中‘花孔雀’在培养40 d后形成愈伤组织,55 d形成不定芽,诱导率可达69.44%;‘黑天鹅’在培养45d后形成愈伤组织,65d形成不定芽,诱导率达到66.67%;最适体细胞胚诱导培养基为MS+2 mg·L~(-1) 6-BA+1 mg·L~(-1) PIC,‘花孔雀’和‘黑天鹅’的诱导率分别达到66.67%和63.89%;最佳不定芽增殖培养基为MS+2 mg·L~(-1) 6-BA+1 mg·L~(-1) NAA+1mg·L~(-1) TDZ,‘花孔雀’和‘黑天鹅’的增殖系数分别达到4.67和3.46;在不添加植物生长调节剂的MS培养基中进行生根培养,30 d后两个品种的生根率均达到100%;将生根培养30 d的小植株转移至室温条件下放置3 d,摘去封口膜再驯化3 d后,移栽至经高温消毒的草炭︰蛭石(体积比)为1︰1的基质中,成活率达到100%。     

苹果离体叶片高效再生不定芽技术研究

苹果不同品种叶片再生不定芽能力不同,嘎拉再生能力最强,其次是乔纳金,富士最难再生。用继代培养３ 年以上、当代培养３０ ～５０ ｄ 的试管苗,取顶部第２ ～４ 叶位幼嫩叶片,远轴面向下接触培养基,再生效率高。诱导叶片再生不定芽适宜培养基为ＭＳ＋ ＴＤＺ１．０ ｍｇ·ｌ－ １（ 富士、乔纳金）或ＢＡ５ ．０ ｍｇ·ｌ－ １ ＋ ＮＡＡ０ ．１ ～０．２ ｍｇ·ｌ－ １ ＋ 蔗糖３５ ．０ ｇ·ｌ－ １ ＋ 琼脂６ ．２ ｇ·ｌ－ １（ 嘎拉） 。     

香玲核桃离体叶片再生体系的建立

为解决核桃种质资源的实验室保存及转基因研究等问题,研究了暗培养时间、叶片放置方式及不同浓度激素组合(BA、NAA、TDZ)对香玲核桃叶片再生的影响。试验结果表明,最适宜的暗培养时间为14d;近轴面放置效果最好;不同组合的激素配比对叶片再生不定芽能力不同,最适宜香玲核桃叶片再生的培养基为:MS+BA0.5mg·L-1+NAA0.5mg·L-1+TDZ1.0mg·L-1+sucrose30g·L-1+agar6g·L-1,在该培养基上,离体叶片不定芽再生率达60%,每叶片平均再生不定芽数超过1.1个。筛选得到了香玲核桃离体叶片的最适分化及生根培养基,对离体再生培养条件进行优化,为香玲核桃的种质资源保存及遗传转化研究奠定了基础。     

四倍体茄子叶片离体培养和植株再生体系的建立

茄子(SolanummelongenaL.)在整个生育期中易受多种病虫害侵袭,特别是黄萎病和青枯病,常给茄子生产带来重大损失。到目前为止仍没有找到行之有效的防治措施和抗病品种。而茄子野生种和近缘野生种中存在高抗黄萎病和青枯病等的珍稀基因,利用这些抗性基因选育高抗的茄子品种是一种     

培养条件对酸枣叶片不定梢再生率的影响

以酸枣无菌苗叶片为外植体,选用基本培养基ＮＮ６９和ＷＰＭ,外源激素包括ＴＤＺ、ＩＡＡ、ＩＢＡ和ＮＡＡ,暗培养时间为２、３、４周。采用正交实验设计方法,研究了基本培养基、外源激素及暗培养时间对叶片不定梢再生的影响。结果表明在附加ＴＤＺ１~３ｍｇ/Ｌ的ＷＰＭ和ＮＮ６９上都可诱导不定梢再生,但ＷＰＭ比ＮＮ６９更有效。暗培养时间对不定梢再生也有影响,暗培养３周比２、４周更有利于提高叶片的不定梢再生率。在附加ＴＤＺ１ｍｇ/Ｌ、ＩＡＡ０．１ｍｇ/Ｌ的ＷＰＭ培养基上,暗培养３周后转到光下培养,获得的不定梢再生率达８７．５%。     

珠美海棠叶片离体培养与植株再生

将珠美海棠试管苗叶片接种于附加NAA0.3 mg·L-1和不同体积分数的6-BA(1.0～6.0 mg·L-1)的MS培养基中进行离体培养,结果表明,6-BA为1.0～3.0 mg·L-1的处理芽再生率为6.7%～31.8%,而根的再生率为26.7%～28.3%.6-BA为4.5和6.0 mg·L-1处理芽再生率分别高...     

秋福红树莓叶片离体再生植株研究

以红树莓品种秋福(Rubus L.cv.Autumn Bliss)离体叶片为外植体,研究适合其再生植株的叶片部位、放置方式、植物生长调节剂以及暗培养时间等条件。结果表明,叶片外植体接种于MS+TDZ2.00mg/L+IAA0.10mg/L的培养基,暗培养2～3周转移至正常光照下培养效果最好,愈伤组织形成率、不定芽再生率和外植体不定芽数分别为100.00%、95.83%和(5.57±0.27)个。将再生芽接种于1/2MS+IBA0.10mg/L的培养基中,35d后生根率达100.00%。再生植株炼苗后移栽,30d时成活率达97.30%,成功地建立了红树莓叶片的离体再生体系。     

合欢离体培养及高效再生体系的建立

以合欢种子和当年生枝条为材料,研究了合欢离体培养及再生体系,探讨了不同激素组合对合欢愈伤组织的产生、分化及生根的影响,筛选出高效产生愈伤组织培养基为MS+BA2mg/L+KT0.5mg/L+NAA0.3mg/L;分化培养基为MS+BA1mg/L+NAA0.1mg/L;有利于生根的培养基为1/2MS+NAA0.5mg/L,以上培养基均加30g/L蔗糖、8g/L琼脂。     

朱顶红离体培养快速繁殖体系及胚状体发生

 以朱顶红鳞茎基部鳞片为外植体, 在MS + NAA 1 mg/L + BA 2 mg/L 培养基上愈伤组织的诱导频率最高, 为93. 3 % , 愈伤组织上可以直接生芽, 生芽频率为90. 0 %。NAA 的诱导效果优于2 ,42D , BA 优于KT。愈靠近鳞茎基部愈容易诱导产生愈伤组织和不定芽。愈伤组织继代培养4 次后有胚状体产生。诱导产生的不定芽和胚状体在MS0 培养基上可以100 %生根。生根后的再生植株移栽到蛭石中, 浇MS 无机盐营养液, 成活率达98 %以上。不生根直接将不定芽移栽到消毒蛭石中, 成活率为85 %以上。     

Effect of insulin on the SGK1 expression and extracellular matrix synthesis in human mesangial cells cultured in high glucose

AIM:To study the effect of insulin on the serum and glucocorticoid-inducible kinase 1 (SGK1) expression and extracellular matrix synthesis in human glomerular mesangial cells (HMC) cultured in high glucose. METHODS:The HMCs were cultured in the presence of 5.5 or 25 mmol/L glucose with or without 100 nmol/L insulin (i.e. NG, HG, NI and HI groups). 4 h latter, expressions of SGK1, insulin receptor substrate-1 (IRS1) and IRS2 in corresponding groups were detected by immunofluorescence or examined by Western blotting. The phosphorylation of IRS1 and IRS2 was measured by immunoprecipitation. 24 h latter, connective tissue growth factor(CTGF) and fibronectin (FN) were also examined by RT-PCR and ELISA, respectively. RESULTS:Compared with NG, the SGK1 protein expression in HG, NI and HI groups was significantly higher (P<0.01). High glucose mainly caused IRS2 protein and its phosphorylation level increase (P<0.01). When treated with 100 nmol/L insulin, IRS1 protein and its phosphorylation in HI group apparently elevated while slight inhibition of IRS2 protein expression and its phosphorylation were observed (HI vs HG, P<0.05). High glucose enhanced the expression of CTGF and FN, and insulin strengthened this effect.CONCLUSION:Insulin and high glucose up-regulate the expression of SGK1 in mesangial cells through different target molecular pathways and ultimately enhance ECM synthesis. The effect of insulin is highly associated with IRS1 signaling cascades.     

Notch1 promotes renal tubulointerstitial fibrosis in renal tissues of diabetic mice by inhibiting autophagy through Akt/mTOR signaling pathway

AIM: To observe the changes of Notch1 expression and autophagy in the renal tissues of diabetic mice, and to explore the regulatory effect of Notch1 on tubulointerstitial fibrosis by inhibiting autophagy in diabetic nephro-pathy. METHODS: The mice were randomly divided into normal control group (db/m mice) and diabetes group (db/db mice), with 8 rats in each group. After 12 weeks of feeding, the mice were sacrificed and the corresponding biochemical indexes were measured. The protein expression of Notch1 in the renal tubular epithelial cells was observed by immunohistochemical staining. The protein levels of Notch1, PTEN, p-Akt (Thr308), Akt, p-mTOR (Ser2448), mTOR, LC3, P62, collagen type Ⅰ (Col-Ⅰ) and collagen type Ⅲ (Col-Ⅲ) were determined by Western blot. RESULTS: Compared with the db/m mice, the blood glucose, glycosylated hemoglobin, serum creatinine, triglyceride and total cholesterol were increased in the db/db mice (P<0.01). Renal tubular epithelial cell vacuolar degeneration, renal tubular expansion and interstitial inflammatory cell infiltration in db/db mouse renal tissues with HE staining were observed. The images of Masson staining showed collagenous fiber-like substance deposition in the glomerular capillaries and renal interstitium, and disarrangement of tubular structure in the renal tissues of db/db mice. The protein expression levels of PTEN and LC3-Ⅱ were decreased (P<0.01 or P<0.05), while the protein levels of Notch1, P62, p-mTOR (Ser2448), p-Akt (Thr308), Col-I and Col-III were increased in the db/db mice as compared with the db/m mice (P<0.01). However, no significant change of total mTOR and Akt proteins between the 2 groups was found. CONCLUSION: Notch1 protein expression was increased, PTEN expression was significantly reduced, Akt/mTOR pathway was activated, autophagy was inhibited, and fibrosis was aggravated in the renal tissues of the diabetic mice.     

Change of ICAM-1 expression in intestine tissue of mice induced by LPS and role of p38 MAPK in its expression

AIM: To study the change of intercellular adhesion molecule-1(ICAM-1) expression in intestine tissues of mice induced by LPS and regulatory effect of p38 mitogen-activated protein kinase(p38 MAPK) on ICAM-1 expression. METHODS: Protein and mRNA of ICAM-1 were measured using Western blotting and RT-PCR respectively in intestine tissue of BALB/c mice treated by lipopolysaccharide(LPS) or LPS plus SB203580, a specific inhibitor of p38 MAPK. RESULTS: Compared with control group, the expression of ICAM-1 protein and mRNA was increased significantly by LPS stimulation in dose- and time-dependent manner. ICAM-1 expression reached peak value at 12-36 h after LPS stimulation. 20.0 mg/kg of LPS could induce the maximum of ICAM-1 expression. Pretreatment of mice with SB203580 for 30 min could inhibit significantly LPS-induced expression of ICAM-1 protein and mRNA expression in mouse intestine tissues. CONCLUSIONS: These data highlight that LPS could up-regulate ICAM-1 protein and mRNA expression in intestine tissue of mice in dose- and time-dependent manner, and p38 MAPK signal pathway plays an important role in ICAM-1 expression induced by LPS. It suggests that inhibition of p38 MAPK might be a useful principle for the prevention and treatment of intestine damage of endotoxic shock.