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1.
橡胶树次生体胚发生及其再生植株遗传稳定性的分子检测   总被引:3,自引:0,他引:3  
以热研7-33-97花药为材料,建立橡胶树次生体胚发生和植株再生体系,主要包括初生花药体胚诱导、次生体胚发生和次生胚植株再生三个步骤.结果表明,来自花药初生异常胚和初生子叶期胚的0.2~0.3 cm胚块均能成功诱导次生体胚发生,并均能获得成熟子叶期胚.以初生异常胚为外植体,每个胚块获得子叶期胚数为0.64个.以初生子叶期胚为外植体连续3次诱导次生体胚发生,随诱导次数增加,体胚发生频率先上升,后保持稳定.2,4-D对次生体胚植株再生具有明显影响.添加4.5 μmol/L和9 μmol/L 2,4-D植株再生培养基中植株再生频率达到85.0%.AYLP检测次生体胚植株遗传稳定结果显示:(1)来自初生异常胚的次生体胚植株60.0%植株DNA发生变异;(2)来自初生子叶期胚的次生体胚植株,随次生体胚发生次数增加,检测到DNA变异植株频率由30.0%升高到52.4%.AFLP聚类分析结果显示,次生体胚植株与热研7-33-97高度相似,相似系数大于0.904.  相似文献   

2.
橡胶树花药培养体胚发生率低及植株再生频率低是制约其组培苗工厂化生产的瓶颈。以热研7-33-97品系花药为外植体,研究橡胶树花药体细胞胚发育过程并对其体细胞胚进行系统分类,同时改良其培养条件。结果表明,完整的体胚发育过程是球形胚-心形胚-鱼雷胚-子叶胚,但有些心形胚不能进一步发育成鱼雷胚,在后继培养中会分化成畸形胚;体胚发生不具同步性,在鱼雷胚旁又长出次生胚。胶树花药体细胞胚共分9种类型,发现胶树花药体胚发生过程中产生连体胚现象,该类胚能正常发育成植株。不同胚状体培养条件对植株再生率的影响说明,暗培养诱导胚状体2个月后再转移至500 lx弱光照条件下培养10~15 d,然后再转移至出苗培养基培养,能够显著提高植株再生率。在诱导胚胎发生培养基中添加不同浓度的稀土,结果表明,稀土具有提高橡胶树花药体胚发生率的效果,其中以添加1 mg/L稀土培养效果最佳。  相似文献   

3.
侵染后抗性胚状体诱导频率低是影响橡胶树遗传转化发展的主要原因,而橡胶树次生体胚发生体系具有体胚增殖效率及植株再生率高的优点。以3个综合性状优良的橡胶树无性系次生体细胞胚状体为受体,研究其对根癌农杆菌的耐侵染能力、GUS基因的瞬时表达情况,探讨不同无性系胚状体对农杆菌转化的反应。结果表明:3个无性系农杆菌污染情况无显著差异,长菌的外植体经无菌水和抗生素水洗涤3 d后即可恢复生长;侵染后每30个胚褐化了1.2~5个胚,占侵染总胚数的4.00%~16.67%,品种间褐化面积无显著差异;热研87-6-62的总蓝斑数显著高于其它2个无性系,平均每个胚染上14.29个蓝斑,热研7-33-97和PR107稍差,但每个胚也能分别染上8.14和8.72个蓝斑。因此3个无性系均可以胚状体作为遗传转化的受体。此结果对利用胚状体为受体改良橡胶树优良无性系奠定良好基础。  相似文献   

4.
牻牛儿基牻牛儿基焦磷酸合成酶(GGPS)和膜结合焦磷酸酶(V-PPase)是橡胶生物合成的重要调控酶.本研究通过双色荧光原位杂交技术对巴西橡胶树GGPS和V-PPase基因进行染色体定位,结果表明:GGPS基因位于巴西橡胶树‘热研7-33-97’4号染色体的短臂上,基因位点距离着丝粒的百分距离为72.39.而V-PPase基因位于巴西橡胶树‘热研7-33-97’10号染色体的短臂上,基因位点到着丝粒的百分距离为25.78.为橡胶树遗传育种及基因工程等提供分子细胞遗传学依据.  相似文献   

5.
巴西橡胶树是一种重要的热带作物。以巴西橡胶树‘热研7-33-97’处于减数分裂四分体时期的幼嫩雄花为试验材料来获取单细胞,利用5%纤维素酶、5%果胶酶、5% β-葡聚糖酶、5%纤维素酶与4%果胶酶混合酶4种酶液,探索酶解四分体胼胝质的最佳方案。结果表明:在37℃条件下酶解1 h,5%纤维素酶与4%果胶酶混合酶的酶解效果最佳,可得到品质良好的小孢子单细胞。通过对拉制针尖细长度、磨针斜口直径以及显微操作分离体系的不断探索与试验,成功建立一套适合巴西橡胶树‘热研7-33-97’小孢子单细胞的显微分离技术体系。小孢子单细胞是减数分裂的直接产物,对小孢子单细胞进行分离并分析可精确解析橡胶树减数分裂过程中基因的重组与交换,确定橡胶树基因组的变异情况。该研究结果将为橡胶树单细胞全基因组测序提供理论与技术支持。  相似文献   

6.
在现有割胶制度下对巴西橡胶树3个品种(热研8-79、热研7-33-97、PR107)的死皮病进行了调查。结果表明:这3个品种的幼龄树发病率和停割率从大到小依次为热研7-33-97(开割4年)> PR107(开割6年)>热研8-79(开割3年),中龄树发病率和停割率从大到小依次为PR107(开割15年)>热研7-33-97(开割8年)>热研8-79(开割14年)。3个品种的中龄橡胶树比幼龄橡胶树的死皮病发病率、停割率和病情指数都有所上升。总体来看,对于幼龄橡胶树而言,肥水条件较好的地块,年干胶株产量高,发病率和停割率低;而中龄树除热研7-33-97表现出幼龄树的趋势外,其它2个品种(热研8-79和PR107)则干胶株产量高,发病率和停割率也高。绝大多数死皮病属割面干涸类型,少部分为褐皮病;褐皮病有一部分是一开始就发生的,多数是由割面干涸发展而来的。在停割至少1年之后,能够恢复排胶的死皮树比例较小,很少超过10%。在3个橡胶树品种中, PR107品种恢复排胶的比例最高,幼龄树和中龄树恢复率分别达8.70%和6.20%。  相似文献   

7.
在橡胶(Hevea brasiliensis)花药组织培养和胚性愈伤长期继代增殖技术的基础上,分别以巴西橡胶热研7-33—97品种的花药脱分化愈伤组织和经继代增殖的花药胚性愈伤组织为转化受体,进行橡胶农杆菌转化体系比较研究。结果表明:胚性愈伤组织是更适宜的转化受体。  相似文献   

8.
为了探寻橡胶树新品种热研7-33-97适宜的割胶制度,在充分挖掘橡胶树产胶潜力的同时又能保证胶树的安全。对热研7-33-97进行1/2阴阳线轮换与单阳线割制的比较。通过5年的田间试验观察,结果表明,在3天1刀的情况下,热研7-33-97优良无性系采用1/2阴阳线轮换割胶制度,其产量比单阳线割制增产10.6%,干含提高0.9%,死皮发病率下降0.5%。  相似文献   

9.
以大规模推广的橡胶树品种热研7-33-97为材料,对橡胶树花药愈伤发生的形态学、组织学进行研究,为胚与非胚性愈伤组织的区分提供试验依据,在橡胶树组织培养和遗传转化中具有重要的意义.结果表明,花药胚与非胚性愈伤组织有着明显差别,前者表现为组织疏松脆性,表面凹凸不平,有较多小球状突起,细胞中含有较大的胞核,细胞形状呈规则小圆球形,细胞排列紧密,细胞体积较小;后者存在两种类型,第一种表现为结构紧密,整块组织硬实,表面较平滑,细胞形态不规则,胞质稀少,部分细胞含有极小的胞核,细胞体积大,第二种表现为组织松软、水渍状,细胞不含有胞核,胞质极稀薄并液泡化,细胞形状不规则,细胞排列较疏松,胞间间隙较大,细胞体积较大.热研7-33-97花药胚性愈伤发生率较低,仅为42%,但分化力强.  相似文献   

10.
橡胶树三倍体种质创制及生物学鉴定   总被引:1,自引:0,他引:1  
以人工诱导的橡胶树三倍体种质为材料,通过形态学、细胞学和分子生物学等方法对其进行鉴定.结果表明:橡胶树通过有性阶段诱导产生的2n配子与正常配子(1n)杂交可获得纯合三倍体,其无性繁殖后代倍性表现稳定,体细胞染色体数为2n=3x=54,三倍体细胞百分率达91.5%;RAPD及过氧化物同工酶分析表明,该橡胶树三倍体与热研7-33-97、RRIM600的亲缘关系较近,在分子水平存在较大的相似性,可判断热研7-33-97、RRIM600为该橡胶树三倍体杂交亲本来源.  相似文献   

11.
Soy protein concentrate (Promosoy R Plus) was mixed with water to form a thick paste and autoclaved at 121°C for 10 min, 30 min, 2 h, or 4 h. Unautoclaved SPC served as a control. Nitrogen solubility measurements and SDS-PAGE analysis indicated that autoclaving resulted in the formation of soy protein aggregates, with a MW of 1 million daltons, held together by non-covalent and disulfide bonds. The 10 min, 30 min, 2 h, and 4 h SPC samples contained 6, 20, 27 and 39% less cysteine, respectively, than the SPC control. No significant differences were found in the PERs, 2.6-2-7, of a casein control, unautoclaved SPC control and 10 min and 30 min autoclaved samples. PERs of the 2 h and 4 h autoclaved samples, 2.0 and 1.9 respectively, were significantly lower than the other four diets. No significant differences were found in the apparent digestibility of the 10 min, 30 min, and 2 h autoclaved samples; the 4 h autoclaved sample however was significantly less digestible. Decreased PERs of autoclaved SPC samples were likely due to (1) crysteine destruction, (2) decreased protein digestibility, and (3) decreased food intake.  相似文献   

12.
植酸广泛存在于禾谷类和豆类等作物种子中,近年来利用诱变技术选育低植酸作物新品种已成研究热点。本研究在不同NaCl浓度处理下,对前期通过60Co-γ辐射获得的9份低植酸小麦突变体材料进行萌发试验。结果表明,0.5%NaCl能促进种子萌发,而1.0%、1.5%NaCl明显抑制种子萌发,在其胁迫下,种子幼苗和主根生长受到抑制。初步筛选到耐盐性较好的低植酸小麦突变体材料4份,分别为lp14、lp49、lp55、lp50。  相似文献   

13.
Some studies have shown that catecholamines and the changes in their levels during and after head trauma can be useful in predicting the outcome in head trauma patients. The goal of this study is to search for a probable relation between urine levels of catecholamines and prognosis in patients with severe head trauma. Fifty four patients with severe head trauma Glasgow Coma Scale (GCS < or = 8) on admission time were recruited in Imam Reza Hospital within one. These patients were included when having no major accompanying trauma in other organs. Twenty four hour urine was collected after admission and levels of metanephrine and nor-metanephrine were measured. The relation between urine levels of these metabolites with final outcome and also with GCS at admission, 24, 48 h and 1 week after admission and discharge time and Glasgow Outcome Scale (GOS) were studied. Fifty two patients, 48 males and 4 females with a mean age of 32.3 +/- 14.7 (3-72) years were included. The main underlying etiologies were motorcycle (46.2%) and car accidents (25%). Diffuse axonal injury, brain contusion and subdural hematoma were three main diagnoses (28.8, 17.3 and 15.4% of the cases, respectively). 19 (36.5%) of the patients expired within the study period. The mean level of metanephrine and normetanephrine in urine were 207.9 +/- 200.5 and 330.2 +/- 218.4 microg in 24 h, respectively. There was no meaningful relation between urine levels of these metabolites and any of GCS and GOS. There was also no meaningful relation between these parameters and final prognosis in patients.  相似文献   

14.
温度预处理对二倍体马铃薯花药培养诱导愈伤的影响   总被引:2,自引:0,他引:2  
以原始二倍体栽培种Solanumphureja-S.stenotomum(PHU-STN)杂种无性系BD10-7和BD29-1的花药为试验材料,研究温度预处理对二倍体马铃薯花药培养过程愈伤组织诱导阶段的影响。预处理采用4℃低温、35℃热激分别进行暗培养24h、48h、72h,预处理后在22℃条件下进行花药培养,以愈伤组织的诱导率为指标探求最佳的预处理方法和水平。结果表明:4℃低温预处理48h后接种花药可明显提高愈伤组织诱导率;两个材料接种后进行热激处理愈伤组织诱导率表现不一致。  相似文献   

15.
为了使EMS诱变在豇豆育种中发挥最佳效果,获得优质的诱变材料,对豇豆品种‘天畅9号’、‘银豇王’进行了3种EMS浓度、3种处理时间的诱导处理,并对当代的种子萌发、幼苗生长、植株主要农艺性状及生化指标的变化进行研究。结果表明:EMS能导致豇豆种子的发芽力和生长速率下降,叶片变小,而种子浸出液电导率、幼苗SOD活性及可溶性蛋白质含量增加;对银豇王的叶绿素含量、一级分枝数、结荚性有增加倾向,而天畅9号表现相反。处理剂量以浓度0.3%EMS、时间3~4 h为宜。  相似文献   

16.

Background

The most common cause of ocular morbidity in developed countries is dry eye, many cases of which are due to lacrimal insufficiency. Dry eye affects approximately 10 million in the United States., most of whom are women. In the U.S. alone, an estimated 2 million Sjögren's syndrome patients have dysfunctional lacrimal glands and severe dry eye, and there is no satisfactory treatment. These patients would benefit if their lacrimal tissue function could be restored.

Methods

The effect of adenovirus-mediated transfer of tumor necrosis factor (TNF)-α inhibitor gene on induced autoimmune dacryoadenitis was evaluated in a rabbit model. Soluble transgene protein was detected in tears by ELISA for 7 days following transduction.

Results

Two weeks after induction of disease with activated lymphocytes, tear production, as determined by Schirmer testing, was reduced by about 40%, while tear film stability, as measured by tear breakup time (BUT), declined by 43%. Adenovirus-mediated gene therapy using AdTNFRp55-Ig given 2 weeks after disease induction, resulted in the return of tear production to normal levels by week 4. In the treated disease group, tear BUT improved significantly by week 4. Rose bengal scores, an indicator of corneal surface defects, increased after disease induction and declined after gene therapy. In the lacrimal gland, the CD4 to CD8 T cell ratio was 4:1 in the disease group compared to 1:2 in the treated group. Infiltration of T cells and CD18+ cells was reduced approximately 50% after gene therapy.

Conclusion

We concluded that therapeutic levels of soluble TNF inhibitor were achieved in the lacrimal gland and on the corneal surface. Anti-inflammatory cytokine gene expression might offer a potential therapeutic modality for the treatment of autoimmune dacryoadenitis, once suitable vectors become available.
  相似文献   

17.
In this study, we synthesized a novel N-halamine precursor, sulfuric acid mono-[2-(4-[4-chloro-6-(2-[4,4- dimethyl-2,5-dioxo-imidazolidin-1-yl]-ethylamino)-[1,3,5]triazin-2-ylamino]-benzenesulfonyl)-ethyl] ester sodium (TB), which contains two reactive groups of monochloro triazine reactive groups and bis-sulphatoethylsolphone reactive groups. The structure of TB is similar to iso-bifunctional group reactive dyes and could be coated on cotton fabrics by covalent bonds through a reactive dyeing process. The cotton coated with TB was characterized by FTIR and SEM. After chlorination, the treated cotton fabrics showed excellent antibacterial efficacy and inactivated all inoculated S. aureus (ATCC 6538) and E. coli O157: H7 (ATCC 43895) within 1 min of contact. Over 85 % of tensile strength retained both in warp and weft directions after treatment and chlorination. Almost 80 % of active chlorine can be regained by treating with household bleach after extensive washing and long time storage.  相似文献   

18.
在辽宁锦州分别采用根瘤菌株S_(4-32)、S_(6-31)、S_(16-4)、S_(13-31)、NC43.3进行覆膜和不覆膜的花生接种栽培试验。花生生长60天不覆膜处理花生结瘤数为16—35个,株平均根瘤干重为18—58mg,而覆膜处理的平均单株根瘤数为70—132个,根瘤干重为119—180mg,覆膜处理的花生植株苗期含氮百分率降低,而总氮积累明显提高,在海花1号品种上,覆膜处理,菌株S_(4-32)具有明显的增效作用,生长60天的平均单株根瘤数、根瘤干重、植株干重和总氮分别达到101个,180.3mg、11.0g,和332.7mg,不覆膜不接菌仅为16.8个,18.7mg、7.3和274mg,前者的荚果产量4224.4公斤/公倾,对照为2955.9公斤/公倾,增产率达42%以上。  相似文献   

19.

Background:

The protein hormone granulocyte colony-stimulating factor (GCSF) stimulates the production of white blood cells and plays an important role in medical treatment of cancer patients.

Methods:

An efficient process was developed for heterologous expression of the human GCSF in E. coli BL21 (DE3). The feeding rate was adjusted to achieve the maximum attainable specific growth rate under critical value. In this method, specific growth rate was maintained at the maximum value of 0.55 h-1 at the beginning of feeding to 0.4 h-1 at the induction time. Recombinant human GCSF (rh-GCSF) was produced as inclusion body. At first, inclusion bodies were released by cell disruption and then washed, solubilized and refolded. Finally, the rh-GCSF was purified by cation exchange chromatography.

Results:

Obviouly, higher specific growth rate decreases process time and consequently increases productivity. The final concentration of biomass and GCSF was achieved 126 g DCW.l-1 and 32.1 g.l-1. Also, the final specific yield (YP/X) and total productivity of rh-GCSF were obtained 254 mg.g-1 DCW and 1.83 g.l-1.h-1, respectively. According to the available data, this is one of the highest YP/X and productivity that has been reported for any human protein which is expressed in E. coli. Recovery yield of purification process was %40 and purity of recombinant protein was over than 99%. The circular dichroism spectra of purified rh-GCSF, Neupogen® and PD-Grastim showed that all proteins have a similar secondary structure.

Conclusion:

Modified exponential feeding strategy for fed-batch cultivation of recombinant E. coli, results in minimum fed-batch duration and maximum productivity. Key Words: Escherichia coli, Granulocyte colony-stimulating factor (GCSF), Process development  相似文献   

20.
The objective of this research was to evaluate the effect of ozone gas and potassium lactate on lipid oxidation and survival of Salmonella typhimurium on fresh pork. A total of 144 samples of fresh pork samples were cut into pieces approximately (8x5x0.6) cm in size, then (0, 2 and 4%) potassium lactate (KL), inoculated with S. typhimurium was applied by spreading a 0.5 mL cell suspension over each sample. The pork samples were then packed in (6"x8") airtight polyethylene bags with a thickness of 87.5 microm and a volume of 3.5 L with and without ozone. Ozone gas was injected into the plastic bags at a rate of 0, 200, 500 and 1,000 mg h(-1) and the samples later stored at 8 degrees C. Thio-barbituric Acid Reactive Substance (TBARS) and microbial loads were determined on days 0, 5, 10 and 15. Data was statistically analyzed using SPSS software and differences among means detected at the 0.5% confident level using the Scheffe's test. Samples treated with 2 and 4% KL had significantly (p<0.01) lower TBARS value than non-treated samples. Combination of ozone and KL showed inhibitory effects on S. typhimurium in samples. S. typhimurium was sensitive to 4% KL with and without ozone. At 1,000 mg h(-1), ozone improved KL inhibitory effect on S. typhimurium. Ozone and KL are potential substances for inhibition of S. typhimurium.  相似文献   

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