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1.
沙田柚遗传转化的探讨   总被引:6,自引:0,他引:6  
采用含nos基因和人工合成柞蚕抗菌肽D基因AP-D基因的根癌农杆菌对沙田柚外植体进行转基因研究,与根癌农杆菌共培养产生的芽,苗经Km敏感性筛选,获得了若干转基因植株,并对这些植株进行了报告基因nos表达的电泳检测,同位素标记、AP-D基因为探针的点印迹杂交,外源AP-D基因的PCR扩增和Southen杂交。结果显示,报告基因nos、抗菌肽D基因已转入沙田柚植株细胞中。  相似文献   

2.
培育苹果转基因植株的研究   总被引:15,自引:2,他引:13  
采用“皇家嘎拉”苹果白化面外植体,通过根癌农杆菌EHA105介导,证明了培养基中存在生长素可促进基因转化,转化效率比对照提高2倍以上。将外植体下EHA101株系在含有生长素的2基中共培养,获得了转基因植株。采用PCR分析和SouthernBlot核酸杂交,确定GUS基因已整合到苹果植株的染色体上。组织化学染色确定了GUS基因在杆株体内的表达。  相似文献   

3.
异戊烯基转移酶基因导入番茄及转基因植株再生   总被引:15,自引:0,他引:15  
通过根癌农杆菌(Agrobacteriens)介导,利用特异启动子Psag12与异戊烯基转移酶(Isopenyl transferase,ipt)基因构建的嵌合基因Psag12-ipt对5个番茄品种进行遗传转化,结果共获得来自31个不同外植体的44株转化再生植株。感染子叶的分化再生及转化植株田间生长正常。对其中部分转化再生植株进行PCR检测证明为转基因植株,并且在田间表现出生长旺盛及抗叶片衰老的特性。  相似文献   

4.
提高苹果基因转化效率的研究   总被引:6,自引:1,他引:5  
采用根瘤农杆菌(Agrobacterium tumefaciens)强株系EHA105(p35sGUS-intron)研究了影响‘皇家嘎拉’苹果(Mains domestica Borkh.)外植体的β-葡萄糖醛酸酶(GUS)基因的瞬时、稳定表达水平和转基因植株的再生。证明在培养基生长素(IBA、NAA)存在的条件下,外植体的GUS基因的瞬时表达水平提高了3-4倍,而共培养两周后稳定表达水平提高2倍以上,产生9.8个GUS愈伤组织表达区域。白化处理促进外植体的基因转化,白化处理的新梢顶端第一节间外植体GUS表达区域比常用的叶片外植体高4倍。在生长素存在的条件下 2%外植体获得了转基因植株。Southern BlotDNA杂交和组织化学染色分析证明GUS基因已整合到苹果的染色体上,并得以表达。  相似文献   

5.
猕猴桃成熟果实中脂氧合酶基因的克隆   总被引:11,自引:0,他引:11  
陈昆松 Ross.  GS 《园艺学报》1998,25(3):230-235
采用PCR扩增方法,从成熟猕猴桃果实组织中克隆到一个824bp的脂氧合酶(LOX)基因片段(pKLC1),该片段含有275个氨基酸组成的开放阅读框架,它与拟南芥菜、黄瓜、豌豆、土豆、水稻、大豆、烟草和番茄的核苷酸同源性为59.0%~74.6%,氨基酸同源性为63.1%~76.3%。Southern杂交表明,pKLC1片段为一低拷贝数基因家族所编码。  相似文献   

6.
以牵牛萌动种胚为受体,用含有35S启动子-Gus基因-ipt基因-Nos基因的根癌农杆菌LBA44043及含有35S启动子-Gus基因-生长素调控基因-Nos基因的根癌农杆菌LBA4404进行转化。通过对转化及对照植株苗期叶的同工酶分析,发现转化植株叶的过氧化物酶同工酶及酯酶同工酶谱带来对照植株有明显差异。转化频率较高。间接验证了外源DNA的导入,说明同工酶分析方法可做为转基因植株早期筛选的方法之  相似文献   

7.
辣椒子叶离体培养和植株再生体系的建立   总被引:50,自引:3,他引:47  
选用辣椒4个栽培品种和2个杂交亲本的子叶进行诱导不定芽分化、生长及生根成苗试验,从中筛选出3个较好的激素配方,建立了辣椒栽培品种的高效快速离体植株再生体系。外植体分化频率可达100%,每外植体平均分化不定芽数10.9个,出苗8 ̄10株。发现添加氨基酸混合物(LY)对不定芽的诱导分化及生长有较强的促进作用。从子叶外植体培养起至再生苗出瓶需50 ̄55d,最快的1批仅44d。  相似文献   

8.
甘蓝显性雄性不育系的选育及其利用   总被引:19,自引:5,他引:14  
方智远  孙培田 《园艺学报》1997,24(3):249-254
从甘蓝原始材料79-399的自然群体中获得雄性不育突变株79-399-3。研究结果表明,该突变株的雄性不育受一显性不育基因CDMs399-3控制。该基因在不同的遗传背景下表现为环境敏感和环境稳定两种类型,且温度是影响不育稳定性的主要因素。环境敏感的显性雄性不育植株通过自交可以得到显性纯合的雄性不育植株。显性纯合不育植株与普通自交系或相应的姊妹系杂交获得了不育株率达到100%、不育度达到或接近100%的不育群体。根据CDMs399-3基因的这一遗传特点,建立了利用显性雄性不育基因配制甘蓝杂交种的新方法:利用组织培养技术保存和繁殖显性纯合雄性不育材料,然后与遗传差异较小的普通自交系或姊妹系杂交得到不育系。选择园艺性状优良、不育性稳定的不育群体作不育系,与配合力优良的自交系杂交,配制出了优良的杂交组合。试验采种结果表明,携带CDMs399-3基因的甘蓝雄性不育系结实性状优良  相似文献   

9.
龙眼rbcL基因结构分析   总被引:2,自引:0,他引:2  
林同香  陈振光 《园艺学报》1999,26(5):291-296
通过Southern杂交等分析,分离和克隆了龙眼叶绿体DNA L145片段,其中含大部分rbcl基因。通过酶切分析制物理图谱,结合亚克隆和PCR扩增,测序分析了龙眼rbcl基因全序列1836bp,其中基因编码为1428bp,编码475个氨基酸和一个终止密码子,rbcl基因的5’上游区长284bp,在基因上游区-191~-220bp范围内有推测的类似原核生物的启动子序列:-10区的TACAAT,-3  相似文献   

10.
甜瓜再生芽高效诱导方法的研究   总被引:24,自引:1,他引:23  
王建设  陈杭 《园艺学报》1999,26(5):339-340
以甜瓜品种伊利沙白为材料,将子叶外植体直接接种在Bordas等诱导体细胞胚性愈伤组织的培养基上,诱导芽的频率高达90%,成芽外植体平均分化出15个芽。  相似文献   

11.
AIM: Although endovascular radiotherapy inhibits neointimal hyperplasia, the exact alterations induced by β-particles irradiation remain to be elucidated. The objective of this study was to investigate the ability and the cellular mechanism of local β-particles emission from 188Re to inhibit vascular smooth muscle cells (SMCs). METHODS: The SMCs in vitro were irradiated by 188Re with single doses of 2.6 Gy-25.8 Gy. The effects of β-particles on SMCs, such as effective irradiate doses, the period of inhibition for SMCs proliferation, the changes of cell proliferation rate and DNA synthesis rate, cell cycle progression and related gene expression, were investigated by cell count, [3H]-TdR incorporation, cell cycle progression analysis, cell viability and immunocytochemistry, respectivecy. RESULTS: β-particles irradiation with dose of 5.2 Gy could inhibit significantly SMCs proliferation. At dose of 20.6 Gy DNA synthesis inhibitory rate was 92%, SMCs proliferation rate was only 3%. Renoval of 188Re did not abolish the inhibitory effects of β-particles on SMCs proliferation. The expression of P53 was up regulation and PCNA was down regulation after irradiation. CONCLUSION: β-particles from 188 Re was significantly effective and permanent in inhibiting SMCs proliferation, and inhibitory effect was in dose-dependet manner ED50was 5 Gy, the best dose to inhibit SMCs proliferation was 20 Gy. β-particles irradiation induced SMCs to occur G0/G1 arrest, damaged the ability of SMCs reproliferation and led to cell clonogenic death. P53 and PCNA had regulatiory effects on SMCs proliferation after β-particles irradiation.  相似文献   

12.
AIM:To study the effect of L-Arg on plasma content of endothelin (ET) and the expression of proto-oncogene c-fos mRNA in the left ventricle of rats with renovascular hypertensive hypertrophy. METHODS: The level of c-fos mRNA were measured by in situ hybridization. The ET in plasma were measured by radioimmunoassay. RESULTS:After eight weeks of treatment with L-Arg, the expression of c-fos decreased markedly (P<0.01). The ET content in plasma also decreased significantly by L-Arg(P<0.01).CONCLUSION: Plasma ET content and the expression of c-fos in the left ventricle of rats with renovascular hypertensive hypertrophy could be decreased by L-Arg administration.  相似文献   

13.
Abstract

Saskatoon berry (Amelanchier alnifolia Nutt., Rosaceae) and blueberry (Vaccinium corymbosum L., Ericaceae) are substantially equivalent in all characteristics that are important to the consumer, including fruit color, shape, size, nutrition, texture, and uses. In addition, both fruits are native to North America and they have practically identical historical uses and known health benefits. Their composition, processing, nutritional value and metabolism, intended uses, and levels of undesirable substances are compared.  相似文献   

14.
The objective of this study was to establish a cryopreservation protocol for hawthorn shoot apices (Crataegus pinnatifida Bge.). Cryopreservation was carried out via encapsulation–dehydration, vitrification, and encapsulation–vitrification on shoot apices excised from in vitro cultures. We began by showing that cold-acclimation enhanced the regrowth of cryopreserved apices from 10.0 to 65.5% in encapsulation–dehydration. We then decided that the encapsulation–dehydration method was an optimal cryopreservation method for hawthorn shoot apices in terms of its high recovery after cryopreservation as well as its ease of use compared with vitrification and encapsulation–vitrification. In encapsulation–dehydration, the protocol leading to optimal regrowth was as follows: after cold-acclimation at 5 °C in the dark for 2 weeks, excised shoot tips were pretreated for 24 h at 25 °C on hormone-free Murashige and Skoog [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15, 473–497] (MS) basal medium with 0.4 mol/L sucrose, then encapsulated and precultured in liquid MS medium with 0.8 mol/L sucrose for 16 h at 25 °C. Precultured beads were dehydrated for 6 h at 25 °C in the dessicator containing 50 g silica gel to a moisture content of 15.3% (fresh-weight basis) before cryostorage for 1 h. In addition, we examined the effect of adding glycerol to both the alginate beads and loading solution to enhance regrowth after cryopreservation in encapsulation–dehydration. In the present study, it was shown that adding 0.5 mol/L glycerol resulted in high regrowth percentages (82.5–90.0%) in four Crataegus species.  相似文献   

15.
AIM: To examine the autoantibody against α1-adrenoceptor and its biologic activities during the development of renal hypertension. METHODS: Renal hypertension of rat was achieved by clipped renal artery, the titre of autoantibody to α1-adrenoceptor was detected using ELISA immunoassay. Furthermore, the biological offects of these autoantibodies on cultured cardiomyocytes were also examined. RESULTS: After two weeks of clipping renal arteries, both the frequency of occurrence and the titre of autoantibodies to cardiac α1-adrenergic receptor were significantly increased as compared with the control of pre-treatment. The increased autoantibodies lasted for several weeks and then automatically decreased gradually to the pre-clipping level at 12 weeks. The biological effects of these autoantibodies displayed an "agonistic-like" activities on the beating frequency of cultured neonatal cardiomyocytes. CONCLUSION: Autoantibodies against α1-adrenoceptor may play a role in the elevation of peripheral vascular resistance and in the development of cardiac hypertrophy in rats with renal hypertension.  相似文献   

16.
AIM: To investigate and screen the sensitive proteins in the formation mechanism of pathological scars by comparing the results of differential proteomic analysis between pathological scars and normal skin.METHODS: Two-dimensional gel electrophoresis was used to detect the protein expression profiles in 8 keloid patients, 8 hypertrophic scar patients and 3 matched normal skin patients.The proteins that showed differential expression of over 4-fold change were cut and analyzed by MALDI-TOF/TOF mass spectrometry.RESULTS: A two-dimensional protein profiling comparison between pathological scars and normal skin was successfully established.On average, 2 978 spots in keloid, 2 975 spots in hypertrophic scar and 3 053 spots in normal skin were identified using gel analysis software.Compared with normal skin, there were totally 36 differentially-expressed proteins in keloid and hypertrophic scar identified from the spots of over 4-fold change, including 16 proteins in both keloid and hypertrophic scar (8 up-regulated and 8 down-regulated), 11 only in keloid (9 up-regulated and 2 down-regulated) and 9 only in hypertrophic scar (4 up-regulated and 5 down-regulated).CONCLUSION: Proteomic analysis can identify the proteins with variance of pathological scars versus normal skin, thus providing probable new clues to reveal the formation mechanism of pathological scars.  相似文献   

17.
多效唑对猕猴桃离体试管苗生长及内源激素的影响   总被引:18,自引:0,他引:18  
多效唑(PP333)处理猕猴桃试管苗,降低了其生长强度;植株体内的GA3、IAA和ZT含量下降,ABA的含量上升,乙烯释放率增加;并且能降低外源的GA3和IAA促进生长的作用,而外源的GA3和IAA又能不同程度地逆转多效唑的抑制作用,使植株恢复生长。  相似文献   

18.
Zusammenfassung Die Leistungsprüfungen wurden im Zeitraum 1997 bis 2003 mit den Unterlagen Gisela 4 und 5, den Klonnummern 195/20 und 497/8 aus der Gisela-Serie sowie Weiroot 10, 13, 53, 72 und 158 durchgeführt. Dabei dienten Sämlinge von P1 (bulgarische Selektion aus Prunus mahaleb) als Kontrolle. Alle Unterlagen waren mit der Sorte Stella veredelt und im Dezember 1996 in der Versuchsanlage der Agraruniversität in Plovdiv, Bulgarien, im Abstand von 6 m×4,5 m gepflanzt worden. Dabei erfolgte ein Pflanzschnitt. Nach Abschluss der natürlichen Kronenentwicklung wurde jedes Jahr ein Winterschnitt vorgenommen. Der Boden wurde durch mechanische Bearbeitung offen gehalten und nach dem 4. Standjahr wurden die Baumstreifen mit Herbiziden behandelt. Die Wasserversorgung erfolgte durch eine dem natürlichen Gefälle folgende Überflutung, allerdings nicht immer zum optimalen Zeitpunkt, da keine eigene Wasserquelle zur Verfügung stand.Basierend auf den Ergebnissen bis zum Anfang des 7. Standjahres können die untersuchten Unterlagen in zwei Gruppen differenziert werden: starkwüchsig—Weiroot 10, P1 und Weiroot 13; mittelstarkwachsend bis schwachwüchsig—Gi 497/8, Gisela 4, Weiroot 53, Weiroot 158, Gi 195/20, Weiroot 72 und Gisela 5. Letztere zeichnete sich durch besondere Schwachwüchsigkeit aus. Die meisten Wurzelschosser bildeten Gisela 4, Weiroot 10 und Weiroot 13. Weiroot 53, Weiroot 72 und Weiroot 158 entwickelten deutlich weniger und P1, Gisela 5, Gi 195/20 sowie Gi 497/8 keine Wurzelschosser. Den frühesten Blühbeginn induzierte Gisela 4. Die anderen Unterlagen führten, in Abhängigkeit von den Temperaturbedingungen des jeweiligen Jahres, zu einer Verspätung der Blüte: P1 und Weiroot 10 um 1–2 Tage; Gi 497/8, Weiroot 13 und Weiroot 158 um 2–4 Tage; Weiroot 72 um 2–7 Tage; Gi 195/20 um 3–6 Tage; Weiroot 53 um 3–8 Tage und Gisela 5 um 3–10 Tage. Die Reifezeit der Früchte war bei den Bäumen auf Gisela 5 im Vergleich zu den anderen Varianten um 2–3 Tage verspätet. Gisela 5, Weiroot 72 und Gisela 4 induzierten bei der aufveredelten Sorte die höchsten Ertragsleistungen, P1 die geringsten. Bei den Bäumen auf Gisela 5 war die Fruchtgröße geringer als bei den anderen Unterlagen. Bäume auf Gisela 5 brauchen intensive Pflege. Nur wenn alle Produktionsfaktoren und kulturtechnischen Maßnahmen optimiert werden, kann das hohe Ertragspotenzial dieser Unterlage ausgeschöpft werden.  相似文献   

19.
Abstract

This review is based partly on complete articles and partly on abstracts. Three of the 60 articles deal with the total uptake of elements in strawberry plant organs in two different strawberry production systems, both considered as optimal concerning amount and balance of elements. The effect on fruit quality may be dramatic if the level of a particular element is outside this range, but there may also be effects initiated by differences within the optimal range of elements. Most articles refer to product oriented quality, but some focus on consumer oriented quality, as discussed by Shewfelt (1999). The discussion here is on a general basis, so one should keep in mind that there are cultivar differences and that specification of nutrition ideally should mirror the needs of a single cultivar, or a group of cultivars with similar requirements. Also, to get a complete understanding of the subject future reviews should embrace a broader access of information including the effect on plant development of individual elements, such as the role of calcium in fruit firmness and its importance in cell wall structure. However, the intention here is to narrow the information to results that suggest a direct connection between nutrient uptake and fruit quality.  相似文献   

20.
AIM:To investigate the effect of metallothionein(MT) on proliferation of rat vascular smooth muscle cells (VSMCs) stimulated by homocysteine and its mechanism. METHODS:VSMCs proliferation was measured by [3-H]-TdR incorporation, mitogen-activated protein kinase(MAPK)activity were determined by immunoprecipitation method, the intracellular contents of MT and malondialdehyde (MDA)were assayed by -hemoglobin saturation method and TBA reaction, respectively, and lactate dehydrogenase (LDH) leakage was measured by NADH oxidation. RESULTS:Hcy(10-6-10-4 mmol/L) stimulated [3-H]-TdR incorporation by the VSMCs in a concentration-dependent manner. Compared with control, [3-H]-TdR incorporation in VSMCs treated with 0.1 mmol/L Hcy was increased by 4.2 fold (P<0.01). Meanwhile, Hcy enhanced MAPK activity, MDA formation and LDH release (P<0.01)in a concentration-dependent manner. Treatment of VSMCs with MT alone did not change above parameters, compared with control. However, MT (10-6-10-4 mol/L)attenuated significantly Hcy-stimulated proliferation of VSMCs (P<0.01)in a concentration-dependent manner. And MT inhibited obviously Hcy-induced activation of MAPK activity, MDA formation and LDH release. Preincubation of VSMCs with 0.5 mmol/L ZnCl2 for 6 h induced an increase cellular MT content by 5.7-fold (P<0.01). The MT-overexpressed VSMCs resisted Hcy-stimulating action on MAPK activity, MDA formation and LDH leakage (P<0.01). CONCLUSION:These results show that MT has an inhibitory effect on Hcy-induced VSMCs proliferation, and that MT could inhibit Hcy-stimulated MAPK activity and lipid peroxidation.  相似文献   

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