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1.
抗生素对高粱茎尖再生的影响及再生体系的建立 总被引:3,自引:0,他引:3
研究羧苄青霉素、头孢霉素、潮霉素和卡那霉素4种抗生素对高粱茎尖再生的影响及再生体系的建立结果表明,MS培养基中添加甘氨酸、乙二胺四乙酸的铁络合物(Fe-EDTA)和抗坏血酸等有机物有利于高粱茎尖的分化与生长。农杆菌介导的高粱茎尖遗传转化过程中羧苄青霉素为最理想抗生素,最佳浓度为250mg/L。并初步确立了高粱茎尖农杆菌介导的遗传转化过程中抑菌和转化体的筛选策略。 相似文献
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抗生素对高粱离体培养反应的影响 总被引:10,自引:1,他引:9
探讨了 5种抗生素对高粱离体培养反应的影响。在 3种抑菌抗生素中 ,羧苄青霉素和头孢霉素的抑菌效果较好。头孢霉素对高梁外植体离体培养的毒性比羧苄青霉素大 ,羧苄青霉素对愈伤组织的生长起促进作用 ,且只在高浓度才对分化有抑制作用。在农杆菌介导高梁遗传转化时 ,选用 2 5 0~ 5 0 0mg L的羧苄青霉素来抑菌是合适的。高梁不同基因型和外植体对潮霉素和卡那霉素的反应表现不同 ,但对潮霉素的反应比卡那霉素敏感。在筛选转化体时 ,以潮霉素较好 ,使用浓度对茎尖和幼胚以 2 5~ 5 0mg L为宜 ,对愈伤组织则以 5 0~ 75mg L为宜。 相似文献
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高粱茎尖再生体系及其遗传转化影响因子的研究 总被引:4,自引:0,他引:4
本文对高粱茎尖再生体系及其转化影响因子进行了研究。不同基因型的茎尖再生频率有很大差异,说明基因型是影响茎尖再生的一个重要因素。在9种不同的激素组合中,以0.25MG/L KT、0.5MG/LBAP和0.25MG/L IAA对茎尖的再生效果最好。在MS培养基的基础上,采用B5培养基维生素并添加200MG/L L-ASN,10MG/L抗坏血酸和100MG/L PVP可提高茎尖再生频率。对农杆菌介导高粱茎尖转化的因子进行了优化,获得较高转化频率的条件为:茎尖预培养时间3D,在OD600值为0.5农杆菌菌液中感染10MIN,共培养3D。 相似文献
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根癌农杆菌介导的葡萄遗传转化体系的优化 总被引:1,自引:0,他引:1
以梅鹿辄葡萄离体叶片为转化材料,对根癌农杆菌介导的遗传转化过程中涉及的几个主要影响因素:抗菌素羧苄青霉素(Carb)的浓度、乙酰丁香酮(AS)的浓度、农杆菌菌液的浓度及卡那霉素筛选压进行优化。结果表明,抗菌素羧苄青霉素在浓度300mg·L-1下即可有效去除农杆菌,且对胚性愈伤组织的分化的影响不大;乙酰丁香酮(AS)的最适浓度为50μm·L-1;农杆菌菌液的浓度控制在OD6000.5~0.7时转化率最高;卡那霉素的筛选浓度控制在100mg·L-1。利用优化的农杆菌介导法进行转化,对再生苗进行PCR检测,结果显示,外源基因已导入了葡萄基因组中。本研究优化了农杆菌介导的葡萄遗传转化影响因素,并获得了转基因植株。 相似文献
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以甘蓝型油菜(Brassica napus L.)品种浙双758子叶柄外植体为受体,建立农杆菌介导转化体系,研究乙酰丁香酮、羧苄青霉素浓度和潮霉素筛选浓度等对农杆菌遗传转化效果的影响。结果发现,羧苄青霉素对农杆菌的抑制效果以500mg/L最佳,且对子叶柄外植体愈伤组织诱导和分化的影响最小,愈伤组织诱导率和芽再生率分别为75.2%和65.1%。5.0mg/L的潮霉素能完全抑制未转化再生植株的生长,使其最终褐化死亡,在此潮霉素筛选浓度下,获得了32株转化再生植株。共培养时培养基中添加100μmol/L AS的芽再生率为3.9%,显著高于未添加AS的芽再生率(2.0%),说明共培养阶段添加酚类物质乙酰丁香酮有利于转化载体T-DNA的转化。部分转化再生植株经PCR和Southern杂交检测呈阳性,病虫害接种试验表明转化植株对菌核病和小菜蛾有较好的抗性。 相似文献
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阔叶猕猴桃抗生素敏感性及遗传转化的研究 总被引:6,自引:0,他引:6
以阔叶猕猴桃(Actinidia latifolia)叶片为外植体,研究了各种抗生素对形态分化和器官形成的影响,并以叶盘为受体,通过根癌农杆菌介导法获得了抗性愈伤组织和抗性芽,组织化学染色表明gus基因已在植物组织中表达。研究结果表明,对不定芽分化来说,头孢霉素(Cef)效果明显好于羧苄青霉素(Carb)。Cef浓度为300mg.L-1时效果最好,再生频率达100%,平均再生芽数达最大为9.76个芽/外植体。生根过程中高浓度的Cef(≥200mg.L-1)明显抑制生根,而Carb对生根没有显著影响。在卡那霉素(Kan)和Carb组合处理中,Carb对生根没有显著影响,但Kan明显抑制生根。随着Kan浓度的升高,生根率和平均根数迅速减少,当浓度升高到50mg.L-1时,生根完全被抑制。因此,农杆菌介导的阔叶猕猴桃遗传转化中愈伤组织和不定芽诱导过程宜选用300mg.L-1Cef来抑制农杆菌生长,Kan筛选的临界浓度为20mg.L-1。在抗性芽生根过程中Kan浓度应该控制在50mg.L-1以内,并改用Carb作为杀菌剂,生根效果会更好。该研究确定了农杆菌介导的阔叶猕猴桃叶片遗传转化中使用抗生素的种类和用量,为通过基因工程对阔叶猕猴桃进行遗传改良奠定了基础。 相似文献
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花生转化和再生研究进展 总被引:13,自引:0,他引:13
许泽永 《农业生物技术学报》2001,9(2):107-111
90年代以来,随着花生遗传转化和再生技术的进步,美国已分别获得抗病和抗虫转基因花生,取得突破性进展。目前,成功的农杆菌介导遗传转化以花生幼苗嫩叶、子叶为外植体,通过卡那霉素筛选,器官发生再生转化植株;基因枪介导的遗传转化是以胚愈伤为转化材料,通过潮霉素筛选转化体胚,再生转化植株。 相似文献
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为建立高效的棕色棉茎尖遗传转化体系,以陆地棉棕彩选1号为材料,采用农杆菌介导茎尖的遗传转化方法,以培养7 d的棕彩选1号幼苗茎尖为受体,设计正交试验对共培养时间、共培养温度、菌液OD600、侵染时间等条件进行优化;此外,还研究了受体茎尖的大小及真空渗透等因素对转化效率的影响,初步建立了棕色棉茎尖农杆菌介导的遗传转化技术体系。结果表明,带有0.5 cm下胚轴的茎尖适合转化,适宜的农杆菌介导遗传转化条件为农杆菌菌液浓度OD600=1.0,侵染时间20 min,共培养时间3 d,共培养温度24℃。此外,真空渗透处理有助于抗性苗的发生。再生植株经在含100 mg·L~(-1)卡那霉素的培养基上筛选,获得102株抗性植株,再进行PCR鉴定和Southern杂交检测,共获得24株转基因植株,转化率为0.4%,且为单拷贝基因插入。本试验建立的优化条件为棕色棉的遗传转化奠定了理论基础。 相似文献
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根癌农杆菌介导的马铃薯转化系统的优化 总被引:1,自引:0,他引:1
以马铃薯品种PB 04的叶片和茎段为外植体,对其再生和农杆菌介导的遗传转化体系进行了研究。分别比较不同转化条件(菌液浓度、感染时间、预培养及共培养时间)对马铃薯遗传转化效率的影响,以及选择压浓度和加入时间对抗性愈伤出芽的影响,结果表明:(1)在玉米素(ZT)2 m g/L,萘乙酸(NAA)0.01 m g/L条件下能有效地提高马铃薯愈伤组织分化率;(2)外植体经过2 d的预培养,在OD600=0.8的农杆菌菌液中侵染10 m in后,抗性愈伤诱导率最高。(3)头孢霉素(C ef)比羧苄青霉素(C ar)更适合用于马铃薯外植体的遗传转化。(4)农杆菌侵染后恢复培养5~7 d再加入卡那霉素(Km)50 m g/L能有效地提高外植体的转化率。 相似文献
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以14个高粱品种为材料,设置5个磷处理水平,对高粱苗期的形态和生理指标进行了研究。结果表明, 1)不同水平磷处理对高粱苗期各形态指标均有极显著影响,其中以0.25 mmol/L作为低磷处理较适合; 2)低磷胁迫对高粱幼苗各形态指标均有不利影响,干重、植株地上部含磷量受低磷的影响较大,可作为耐低磷高粱品种的筛选指标; 3)不同高粱品种对低磷的耐性存在明显的基因型差异, 聚类分析显示八月齐属相对耐低磷性强的品种,农858属低磷敏感型品种; 4)低磷下叶绿素和可溶性蛋白含量下降,丙二醛(MDA)含量及超氧化物歧化酶(SOD)、过氧化物酶(POD)活性升高,除可溶性蛋白外,叶绿素、 MDA、 SOD和POD活性均表现为耐低磷性强的相对变化率较小,耐低磷性弱的相对变化率较大。 相似文献
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Primary determinants of crop production in arid/semiarid regions are lack of moisture and infertility, especially phosphorus (P) deficiency or unavailability. The effects of P and water stress (WS) levels on shoot and root dry matter (DM), leaf area, root volume, total root length, and shoot and root P concentrations and contents were determined in two bean [Phaseolus acutifolius Gray, cv ‘Tepary #21’ ("drought‐resistant") and P. vulgaris L., cv “Emerson’ ("drought‐sensitive")] and two sorghum [Sorghum bicolor (L.) Moench, cv SA7078 ("drought‐resistant") and ‘Redlan’ ("drought‐sensitive")] cultivars grown in nutrient solution. Plants were grown with different levels of P (20 and 100 μM for bean and 20, 80, and 160 μM for sorghum) when seedlings were transferred to nutrient solution, and WS levels of 0, 13.8, and 1 6.4% polyethylene glycol (PEG‐8000) introduced after plants had grown in solution 23 days (bean) and 31 days (sorghum). All growth traits were lower when bean and sorghum plants were grown with WS and low P. Growth traits were higher in cultivars grown with high compared to low P regardless of WS. Root P concentration and content and shoot content, but not shoot P concentration, were lower when bean plants were grown with WS compared to without WS. Tepary #21 bean had higher shoot DM, leaf area, total root length, and shoot P concentration than Emerson when plants were grown with WS at each level of P. Sorghum shoot and root P concentrations were higher as P level increased regardless of WS, and WS had little effect on shoot P concentration, but root P concentration was higher. Contents of P were similar for SA7078 and Redlan regardless of P or WS treatment, but SA7078 had greater P contents than Redlan over all P and WS treatments. “Drought‐resistant”; cultivars generally had better growth traits, especially total and specific root lengths, than “drought‐sensitive”; cultivars. 相似文献
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A. Promkhambut C. Akkasaeng A. Younger 《Acta Agriculturae Scandinavica, Section B - Plant Soil Science》2013,63(4):345-355
Abstract Understanding the responses of sweet sorghum to flooding and the characters associated with flooding tolerance may be a useful strategy for pre-rice production and help meet demand for biofuel feedstock. Three sweet sorghum genotypes (Bailey, Keller and Wray) and five flooding treatments including non-flooding control, continuous flooding extended from 30, 45, 60 and 75 days after emergence to harvest were conducted under greenhouse conditions. Flooding decreased leaf dry weight (22–60%), leaf area (10–70%), number of node per stalk (1–5%), shoot dry weight (5–20%) and stalk yield (2–22%) with highest reduction in 30 days after emergence flooding treatment. Flooding later than 30 days after emergence did not significantly affect shoot growth, yield and yield components. Brix value, sucrose content and total sugar content were not significantly affected. All studied cultivars had similar shoot growth response. Flooding induced development of roots in water; root length, root dry weight, nodal root and lateral root number and interconnection of aerenchyma spaces from roots in flooded soil to stalk base above water level but suppressed root growth in flooded soil. The acclimation traits were highest in Keller, flooding from 30 days after emergence but there was a lack of root development in 75 days after emergence flooding treatments. These findings indicate the effect of waterlogging on sweet sorghum growth and yield strongly depends on the growth stage at which it occurs. There were genetic variations in root morphological and anatomical responses to flooding of sweet sorghum. The development of nodal and lateral roots and aerenchyma formation from flooded plant parts to stalk bases above water level may distribute to flooding tolerance in sweet sorghum. Based on the results, a flood-free period of at least 30 days after emergence is required to sustain yield of pre-rice sweet sorghum and early planting is highly recommended. 相似文献
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S. Appa Rao K. E. Prasada Rao M. H. Mengesha V. Gopal Reddy 《Genetic Resources and Crop Evolution》1996,43(6):559-567
Summary Of the 5000 sorghum (Sorghum bicolor L. Monench) germplasm accessions assembled at ICRISAT Asia Center (IAC), over 4000 accessions from 11 major sorghum growing states in India were evaluated for morphological and agronomical characters at IAC during rainy and postrainy seasons.Considerable diversity was observed for all the characters studied. Diversity was more among states than within states. Variation among the seasons was also considerable. Days to flowering ranged from 42 to 129 days during postrainy and 33 to 180 days during the rainy season indicating their day length sensitivity. Plant height ranged from 65 to 330 cm during postrainy and 75 to 655 cm during rainy season. Panicle length ranged from 5 cm to 52 cm and panicle width from 1 to 51 cm. Erect and compact panicles are more frequent although all possible panicle shapes and compactness were found. In general, sorghums from India produce medium to large, lustrous grains in which the subcoat (testa) is absent. All the 5 basic and 10 intermediate sorghum races were found in India though durra or half-durra predominate. Racial diversity is maximal in the state of Andhra Pradesh followed by Maharashatra state. In the world collection a majority of the sources of resistance to shoot fly, stem borer, and striga are from India. Based on diversity for morphological and agronomical characters, India could be considered as the secondary center of diversity. Sorghum germplasm from India is useful for good quality large grain. 相似文献
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连作对高粱生长及根区土壤环境的影响 总被引:2,自引:0,他引:2
以玉米-高粱轮作为对照,研究连作对高粱生长、产量的影响及根区土壤中酶活性和微生物的变化。结果表明:高粱连作3年对产量的抑制开始显现。与轮作相比,连作3年高粱的株高、茎粗、叶面积、生物量显著降低,连作4年更为明显;连作4年高粱根系生长也明显受到影响,0~40 cm深度范围内根系的生物量、根表面积和根体积分别仅为轮作的61.5%、84.4%、73.8%。连作4年增加了土壤中可培养真菌数量,在拔节期和灌浆期分别是轮作的1.9、1.3倍,而对细菌和放线菌的影响没有明显规律;连作显著增加土壤中过氧化氢酶、蔗糖酶的活性,分别比轮作增加了14.7%、17.2%。由此可见,连作不仅抑制高粱植株的生长,并对土壤中微生物区系组成和酶活性产生显著影响。 相似文献
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Associations between vesicular‐arbuscular mycorrhizal (VAM) fungi and manganese (Mn) nutrition/toxicity are not clear. This study was conducted to determine the effects of excess levels of Mn on mineral nutrient uptake in shoots and roots of mycorrhizal (+VAM) and non‐mycorrhizal (‐VAM) sorghum [Sorghum bicolor (L) Moench, cv. NB9040]. Plants colonized with and without two VAM isolates [Glomus intraradices UT143–2 (UT1 43) and Gl. etunicatum UT316A‐2 (UT316)] were grown in sand irrigated with nutrient solution at pH 4.8 containing 0, 270, 540, and 1080 μM of added Mn (as manganese chloride) above the basal solution (18 μM). Shoot and root dry matter followed the sequence of UT316 > UT143 > ‐VAM, and shoots had greater differences than roots. Shoot and root concentrations and contents of Mn, phosphorus (P), sulfur (S), potassium (K), calcium (Ca), magnesium (Mg), iron (Fe), zinc (Zn), and copper (Cu were determined. The +VAM plants generally had higher mineral nutrient concentrations and contents than ‐VAM plants, although ‐VAM plants had higher concentrations and contents of some minerals than +VAM plants at some Mn levels. Plants colonized with UT143 had higher concentrations of shoot P, Ca, Zn, and Cu and higher root Mg, Zn, and Cu than UT316 colonized plants, while UT316 colonized plants had higher shoot and root K concentrations than UT143 colonized plants. These results showed that VAM isolates differ in enhancement of mineral nutrient uptake by sorghum. 相似文献