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1.
茶薪菇原生质体制备及诱变的研究   总被引:8,自引:0,他引:8  
对茶薪菇原生质体的分离及再生进行研究 ,结果表明以溶壁酶的去壁效果最好 ,不同酶组合可提高去壁效果 ,酶解液在 2 0 %时原生质体释放量很高。酶解适宜温度为 30℃ ,培养 4~ 5d的菌丝酶解 3h释放的原生质体最多。以甘露醇作为渗透压稳定剂 ,原生质体的释放量及再生率均较高 ,再生率达到 5 %左右 ,经紫外线照射 2 0s,致死率即达 70 %以上  相似文献   

2.
厚环乳牛肝的原生质体分离与再生   总被引:2,自引:0,他引:2  
探讨了外生菌根菌耳环乳牛肝(Suillusgrevillei)的原生质体制备及再生条件。用MYG培养基培养菌丝7天、宋用1.5%Lywallzyme,以0.6mol甘露醇为稳渗剂,在酶解液pN值5.5,温度31℃,酶解4hr的条件下,每0.1g湿菌丝的原生质体产量这6×10 ̄6/ml。用SO培养基培养菌丝能提高原生质体产量,但再生率下降。综合产量和再生率考虑以MYG培养菌丝效果较好。  相似文献   

3.
灵芝原生质体形成与再生的研究   总被引:1,自引:1,他引:0  
该文通过对原生质体形成与再生条件如酶类、渗透压稳定剂、pH、培养基、菌龄、酶解温度和时间等因素的研究表明,以D培养基、葡萄糖为渗透压稳定剂,酶液浓度为15%Lyw+02%Lys,pH45~55,培养菌龄为72~96h,酶解温度为20~25℃条件下,酶解60~90min,其破壁效果为最佳,原生质体形成和再生率都最高。  相似文献   

4.
糙皮侧耳原生质体制备与再生条件   总被引:4,自引:3,他引:1  
原生质体制备是通过体杂交或基因转导进行种质改良的基础。探讨了糙皮侧耳原生质体制备及再生条件。用MYG培养菌丝,以甘露醇为稳渗剂,用1.5%溶壁酶Lywallzyme进行酶解脱壁,在菌龄7天,酶液pHWFHG4.5,温度28℃,酶解4hr的条件下原生质体产量最高(20×10^6/ml,0.1g湿菌丝),其再生率大于8%,菌丝培养前对接种物进行予切碎处理可进一步使产量提高约2倍,而再生率不并显著下降。  相似文献   

5.
在适宜培养条件下,振荡培养4-5d的鸡Zong菌和粗粗柄鸡Zong菌幼嫩菌丝,经过滤洗涤收信,以0.6mol/LKCl为渗透压稳定剂,用1%纤维素酶加0.5%蜗牛酶的混合酶液处理,于28℃酶解3-4h,可获得大量原生质体,产量达2.82*10^6-3.24*10^6/mL。但所形的原生质体再生能力较差,再生率仅0.6%-1.2%。本文还详细研究了菌龄,酶系统,酶解温度和时间等多种因素对这种两鸡Zo  相似文献   

6.
灵芝原生质体制备与再生条件的研究   总被引:4,自引:1,他引:3  
本文研究了不同酶浓度、酶解温度、酶解时间、渗透压稳定剂及菌龄等对灵芝菌丝原生质体制备与再生的影响,结果表明,菌龄为60h的菌丝,采用浓度为2%的溶壁酶以0.6M的甘露醇作渗透压稳定剂于pH6.5,30℃下酶解2h,当酶解液中湿菌体含量对0.1g/mL时,其原生质体数可达2^*10^7个/mL,且再生率达8.5%。  相似文献   

7.
研究了不同酶种类、酶浓度、酶解温度和时间、渗透压稳定剂、pH值及菌龄等因素对云芝原生质体制备以及不同再生培养基对云芝原生质体再生的影响。结果表明,菌龄3d的云芝菌丝体用3%溶壁酶,以pH5.5,0.4mol/L的KCl为稳定剂,30℃酶解5h获得的原生质体产量最高;用RMl再生培养基培养,原生质体再生率最高。  相似文献   

8.
银耳菌丝原生质体分离和再生研究   总被引:3,自引:0,他引:3  
以菌株Tr01为材料,研究了不同菌龄、溶壁酶浓度、酶解温度、稳渗剂对银耳工力丝在的原生质体产量及不同稳渗剂对原生质再生的影响。结果表明,原生质体分离的最佳条件为:菌丝菌龄9天,酶液浓度2%,稳渗剂0.4mol/L硫酸镁,在34℃下酶解4h,原生质体以0.4mol/L甘露醇作稳渗剂再生率较高,为1.13%。  相似文献   

9.
灵芝原生质体的制备与再生研究   总被引:1,自引:0,他引:1  
以泰山灵芝为试材,研究了菌龄、酶浓度、酶解时间、酶解温度、酶解pH、渗透压稳定剂对灵芝原生质体制备和再生的影响.结果表明:菌龄为7d,以甘露醇为渗透压稳定剂,采用酶浓度2.0%的溶壁酶,pH为6.0,31℃条件下酶解2.5h为灵芝原生质体制备的最适条件;并在原生质体再生固体培养基(以0.6 mol/L蔗糖为渗透压稳定剂)上,原生质体的再生率最高.  相似文献   

10.
马铃薯花粉原生质体分离的研究   总被引:3,自引:0,他引:3  
王蒂  司怀军  王清 《园艺学报》1999,26(5):323-326
以马铃薯减数分裂二分体和四分体时期的小孢子为材料,对原生质体分离的主要影响因素进行了研究。结果表明,1%蜗牛酶和0.5%崩溃酶的混合酶液游离效果最好,原生质体的最高得率达65.4%。渗透压调解剂以蔗糖最好,甘露醇次之,最适浓度分别为16%和18%。0.1%荧光增白剂检测表明脱壁完全,FDA荧光检测表明花粉原生质体具有生活力。  相似文献   

11.
12.
AIM: To investigate the linkage between the polymorphism of -109 and Glu237 in the high-affinity IgE receptorβ(FcεRⅠβ) gene and susceptibilty of allergic asthma in adults in a Chinese population. METHODS: Allergic asthma sample in adult and age-and sex-matched control were studied. A-109C/T and a coding variant Glu237Gly in FcεRⅠβ were detected with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: (1)The genotype frequencies were 0.415 for -109T/T, 0.491 for-109T/C and 0.094 for -109C/C in allergic asthma in adults. No significant difference in the distribution of-109C/T polymorphism was found between allergic asthma adult subjects and healthy control subjects. However, a homozygosity for the-109T allele was associated with increased total plasma IgE levels in subjects with allergic asthma (F=7.213, P<0.01). (2)The allele frequency of Gly237 in the patients and control was 0.245 and 0.142, respectively. There was a significant association between Gly/Gly genotype and allergic asthma in adults among allergic asthma patients. Gly237 was significantly associated with high IgE. CONCLUSIONS: It suggests that Gly237 vaviant of the FcεRⅠβ gene is involved in the development of allergic asthma. The-109C/T and Glu237Gly polymorphisms are two of the genetic factors identified, which affects total plasma IgE levels in adult allergic asthma in Han of Hubei province.  相似文献   

13.
AIM: To investigate the possible role of endothelin(ET-1) in asthma pathogenesis and the effect of atrial natriuretic factor (ANF) on changes of ET-1. METHODS: Measuring the contents of endothelin-1(ET-1),atrial natriuretic factor(ANF),cGMP in plasma and bronchoalveolar lavage fluid (BALF) of guinea pigs. RESULTS: The contents of ET-1, ANF and cGMP in asthma group were higher than that of the control group; There was a significant negative correlation between ET-1 and ANF( r=-0.638,P <0.05) in plasma of the asthma group, and a significantly negative correlation between ET-1 and ANF( r=-0.921,P <0.01) in BALF of the asthma group. There was a significantly positive correlation between ANF and cGMP( r=0.848,P <0.01) in plasma of the asthma group,and a significantly positive correlation between ANF and cGMP ( r=0.831,P <0.01) in BALF of the asthma group. The levels ET-1 in the asthma+rat ANF(rANF) group were lower than those in the asthma group,and the levels of cGMP in the asthma+rat ANF(rANF) group were higher than those in the asthma group after ceasing to infuse rANF for guinea pigs for 30 minutes.CONCLUSION: ET-1 might play an important role in the pathogenesis of asthma.ANF might inhibit production of ET-1.  相似文献   

14.
AIM:To investigate the mechanism of interleukin-1β (IL-1β) promoting the transformation of naïve T cells into Th22 cells and the correlation of its peripheral blood expression in non-small cell lung cancer patients. METHODS:CD4+ naïve T cell magnetic bead sorting kit was used to isolate the peripheral blood mononuclear T cells from healthy people. Transforming growth factor-β (TGF-β) and IL-2 were added to promote differentiation and proliferation. IL-1β was used to induce differentiation into Th22 cells. The proportion of CD4+ IL-22+ T cells was analyzed by flow cytometry, and the expression of IL-22 was detected by ELISA. We selected 60 cases of non-small cell lung cancer patients in our hospital, including 18 in I phase, 20 in Ⅱ phase, 13 in Ⅲ phase and 9 in IV phase, as well as 25 healthy persons. The proportion of Th22 (CD4+ IL-22+) cells in peripheral blood was detected by flow cytometry, and the serum levels of IL-1β and IL-22 were measured by ELISA. RESULTS:IL-1β induced the transformation of naïve T cells into Th22 cells and promoted the secretion of IL-22 (P<0.05). The proportion of Th22 cells and the IL-22 and IL-1β levels in peripheral blood of the patients with non-small cell lung cancer were higher than those in healthy subjects, and correlated with the clinical stage. CONCLUSION:IL-1β induces the differentiation of Th22 cells and the expression of IL-22. The levels of IL-1β and IL-22 are related to the progression of non-small cell lung cancer, which may be involved in immunosuppression and promote the occurrence of non-small cell lung cancer.  相似文献   

15.
AIM:To observe changes of beta-adrenoceptor (β-AR) and its relativity to polymorphonulear leukocytes (PMN) and acute lung injury (ALI) in rats. METHODS:ALI modle in rat was established by intravonous injection of E. Coli endotoxin (ET). β-AR was measured by radioligand binding assay with -dihydroalprenlol.RESULTS:(1)Maxmal capacity of β-AR (Bmax) in both lung tissue and PMN decreased significantly at the lst, 4th and 6th hours after injection of ET. (2)No relativity was found between β-AR Bmax changes of lung tissue and β-AR Bmax changes of PMN. CONCLUSIONS:(1) Decrease of density of β-AR in lung tissue and PMN may played a role in the development of ALI. (2) The β-AR of PMN in circular blood can not be used as a relative index of β-AR changes of lung tissue in rats with ALI.  相似文献   

16.
AIM: To explore the role of plasma β-endorphin(β-EP) in the suppression of cellular immunity following trauma-hemorrhagic shock(T-HS). METHODS: ① Wistar rats with T-HS were sacrificed at 0 h, 1 h, 3 h, 6 h, 12 h, 24 h after T-HS. Plasma sample was collected and β-EP levels in plasma was measured. Rats with sham-operated were served as the controls. ②In in vitro experiment, splenic cells were isolated and mixed from four normal rats and cocultured with shock plasma (SP) or SP +β-EP antiserum. ConA-induced splenic cell proliferation, IL-2 production, IL-2R expression were examined. RESULTS: ① Levels of plasma β-EP elevated remarkably after T-HS immediately, peaked at 1h , showed decreasing tendency and restored normal 24 h after T-HS. ② Shock plasma significantly suppressed ConA-induced splenic cell function. Levels of plasma β-EP were negatively correlated with spenic cell proliferation and IL-2 production and IL-2R expression. Compared with SP group, splenic cell function elevated markedly in SP + β-EP antiserum group, but still lower than that in controls. CONCLUSION: The elevated plasma β-EP following T-HS was involved in the suppression of cellular immunity.  相似文献   

17.
AIM: To investigate the effects and mechanisms of irbesartan, one of the angiotensin Ⅱreceptor blockers, on kidney function in diabetic rats. METHODS: Forty adult male Wistar rats were randomly divided into four groups: control group, diabetes group, irbesartan group and captopril group. At the end of 12 weeks, the rats were sacrificed. Urine volume, body weight, kidney weight/body weight, plasma, glucose, glycosylated hemoglobin (HbA1c), urinary β2-microglobulin (β2-MG) excretion, urinary albumin excretion rate (UAR), creatinine clearance (Ccr) were measured. Nitric oxide (NO) and endothelin-1 (ET-1) levels in plasma, urinary and renal tissues were determined. RESULTS: Urine volume, kidney weight/body weight, plasma glucose, HbA1C, UAR, Ccr, urinary β2-MG excretion, NO and ET-1 levels of urinary, blood and renal tissue in diabetic rats were significantly higher than those of normal controls ( P<0.01). UAR, Ccr, urinary β2-MG excretion, ET-1 and NO levels of urinary and renal tissue in rats of irbesartan and captopril groups were significantly lower than those of DM rats ( P<0.01). There were positive relationships among the levels of plasma, urinary, renal tissue ET-1, NO and UAR, Ccr and urinary β2-MG excretion. CONCLUSION: Irbesartan could prevent from the injury of renal function in STZ-induced diabetic rats. And it maybe one of the most importan mechanisms that irbesartan could reduce the NO and ET-1 levels in STZ-induced diabetic rats.  相似文献   

18.
AIM: To clarify the relationship between expression of leukocyte iNOS-mRNA and pancreatic islet function in the diabetic rats induced by streptozocin (STZ). METHODS: Wistar rats were randomly divided into the control group (n=10) and the diabetic group (n=15). Expression of iNOS-mRNA in the peripheral blood leukocyte, liver and lung were detected with in situ hybridization and the blood sugar and insulin were also measured. RESULTS: It showed that the blood glucose content increased from (8.95±1.80) to (22.84±4.90) mmol·L-1, however, the plasma insulin content decreased from (81.76±2.12) to (58.92±18.20) mU·L-1 at the third day after the β cell was disfunctioned by STZ injection. No expression of leukocyte iNOS-mRNA in normal rat was detected. The percent rate of positive cells were significantly increased in the rats with diabetes. CONCLUSION: The expression of leukocyte iNOS-mRNA is positively related to the damage of β cells caused by STZ.  相似文献   

19.
AIM: To explore the effect of Wnt/β-catenin signaling pathway in airway smooth muscle cells (ASMC) on asthmatic airway remodeling.METHODS: The asthmatic airway remodeling model in rats was established and the ASMC was isolated and cultured. The protein expression of β-catenin, glycogen synthase kinase-3β (GSK-3β), c-Myc and cyclin D1 in the ASMC was determined by Western blot. After depressing the interaction between β-catenin and p300/CBP, the cell activity was measured by CCK-8 assay and the change of cell cycle distribution was analyzed by flow cytometry. Meanwhile, the protein expression of c-Myc and cyclin D1 in the ASMC was determined by Western blot after inhibiting P38 mitogen-activated protein kinase (MAPK) activity.RESULTS: The protein levels of β-catenin, c-Myc and cyclin D1 were significantly increased in asthma group while the protein level of GSK-3β was decreased in the same group (P<0.05). After depressing the interaction between β-catenin and p300/CBP, the cell activity of ASMC was decreased in asthma group compared with control group (P<0.05), and the change of the cell cycle distribution in asthma group was also more obvious (P<0.05). After inhibiting P38 MAPK activity, the protein levels of c-Myc and cyclin D1 were all decreased compared with control group in ASMC asthma and control rats (P<0.05).CONCLUSION: Wnt/β-catenin signaling pathway may participates in airway remodeling in asthma by increasing the protein expression of c-Myc and cyclin D1, reacting with the P38 MAPK signaling pathway and regulating the growth of ASMC.  相似文献   

20.
AIM:To study the effect of IFN-γ inhalation on the anti-infection ability of the lungs in the immunocompromised host. METHODS:The immunological factors in the immunocompromised rats and the immunocompromised rats administrated IFN-γ via aerosol were investigated after 1, 3, 7 days when they were injected Candida albicans via tracheal. The Canidda albicans count of the left lung was also determined after 7 days when injecting pathogen. RESULTS:The Canidda albicans count of the left lung in IFN-γ group was significantly less than that of control group. The phagocyting and bactericidal percentages, Ia antigen expression percentages, the levels of TNF-α, IL-1β and IL-6 in the culture supernatant of the AM, the activity of IFN-γ and TNF-α in BALF (except the TNF-α on 7 th day) in IFN-γ group were markedly higher than those in control group. The expression of IFN-γ and IL-1β pulmonary tissues in IFN-γ group was higher than that in control group. The expression of TNF-α in IFN-γ group was less than that in control group. The expression of IL-6 was no changes between two groups. The levels of IFN-γ, IL-1β and IL-6 in the blood (except IL-1β on 3 rd day), and the killing ability of the lymphocytes in blood had no difference between two groups. CONCLUSION:Administration of IFN-γ via aerosol obviously enhanced the anti-infection ability of the lungs in the immunocompromised host, but has no influence on the whole body cellular immunity.  相似文献   

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