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1.
Summary In earlier work on improvement of persistance in forage legumes, we selected genotypes from highly productive cultivars of alfalfa, Algonquin and Apica (Euphytica 45: 105–112, 1990) and cv. Florex red clover (Plant Cell Reports 8: 395–398, 1989) capable of in vitro regeneration from callus and cell culture. The alfalfa germplasm and its F1 progeny as well as an F2 red clover population were tested for cold stress tolerance. Plantlets were hardened in culture tubes at 2 or 5°C, 8h photoperiod, for at least four weeks and then subjected to freezing temperatures, –16 or –10°C for alfalfa and red clover, respectively. Survival of regenerative genotypes was significantly higher than of the non-regenerative ones in both species. A strong oositive correlation (r=0.78) between the regenerative trait and plant survival was found in alfalfa. The experiments indicate that in vitro selection for regenerative trait may improve cold stress tolerance of alfalfa and red clover.  相似文献   

2.
Anther culture in connection with induced mutations for rice improvement   总被引:2,自引:0,他引:2  
Doubled haploids have long been recognized as a valuable tool in plant breeding since it not only offers the quickest method of advancing heterozygous breeding lines to homozygosity, but also increases the selection efficiency over conventional procedures due to better discrimination between genotypes within any one generation. Ten cultivars of japonica rice and nine cultivars of indica rice were evaluated for androgenic response. Various doses (10–50 Gy) of gamma rays were applied to investigate the effect of radiation on callus formation, green plant regeneration and the frequency of selected doubled haploid mutants. Similarly, the effects of colchicine concentration (10–200 mg/l) on callus induction, regeneration and fertility of green plants were observed. It was demonstrated that the dose of 20 Gy gamma rays and 30 mg/l concentration of colchicine have significant stimulation effect on regeneration of green plants from rice anther culture. The high frequency of observed doubled haploid mutants indicates that anther culture applied in connection with gamma rays is an effective way to improve rice cultivars. This revised version was published online in August 2006 with corrections to the Cover Date.  相似文献   

3.
In a screening experiment with 215 different winter cultivars of Triticum aestivum L. for response in anther culture, pollen embryos or callus were obtained from 200 different cultivars and green plants from 93 different cultivars. On average, from the whole material, 1.3 green plants were obtained per 100 anthers cultured. Variance components estimated from replicates with selected parts of the plant material indicated that for the formation of pollen embryos from anthers, interactions between genotypes and replications were dominating, accounting for 45 to 50 per cent of the variation. Main effects from genotypes were less prominent, accounting for 25 to 30 per cent of the variation, and the replications showed relatively little effect, accounting, for about 4 to 12 per cent of total variation. Regeneration of plants from pollen embryos or callus was not significantly influenced by either genotype or replicates. The frequencies of plants regenerated being green, however, were influenced from both genotype and environments contributing 42.5 and 34.9 per cent of the total variation respectively, while only minor interactions between genotype and environments were indicated for this character.  相似文献   

4.
Plant regeneration in sweet potato (Ipomoea batatas L., Convolvulaceae)   总被引:1,自引:0,他引:1  
The application of new techniques for improvement of sweet potato crops, particularly including the exploitation of somaclonal variation, gene transfer by genetic transformation and somatic hybridization, requires the control of plant regeneration from tissue cultures. Shoots can easily be regenerated from explants of stems, petioles, leaves and roots, while callus cultures do not produce any shoots. The potential of somatic embryogenesis and plant regeneration via embryogenesis was evaluated for 10 cultivars of sweet potato. Protocols for plant regeneration from cultured protoplasts have also been developed. Since mesophyll was resistant to enzyme digestion, fragments of stems and petioles, callus and cell suspensions were used as source of protoplasts of sweet potato. Series of transfers of protoplast-derived calluses, particularly those which had been obtained from in vitro plants, to media containing a high level of zeatin resulted in successful formation of shoots in only two sweet potato cultivars. In addition, the embryogenic potential was irreversibly lost through protoplast culture, since protoplasts isolated from embryogenic cell suspensions developed into non-embryogenic callus. Consequently, an alternative protocol is being successfully developed to improve plant regeneration from cultured protoplasts of sweet potato, involving first root formation from which shoots can then be regenerated. Preliminary evaluation in field conditions in Gabon revealed that plants regenerated from cultured protoplasts exhibited a great genetic variability in their growth and tuber formation in particular. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

5.
Summary Calli were obtained from mesocotyl explants of eight varieties of pearl millet using MS medium supplemented with 2,4-D. Variable frequencies of callus initiation were observed among the varieties. Plantlets were regenerated from the calli by transferring on to the medium devoid of auxins. Incorporation of coconut milk, Kinetin, IAA or NAA did not enhance either the callus initiation or growth rate or plantlet regeneration. The eight genotypes differed among themselves in callus growth rate and frequency of shoot bud production.  相似文献   

6.
Plant regeneration through callus initiation from mature embryo of Triticum   总被引:3,自引:0,他引:3  
R. M. Bi    M. Kou    L. G. Chen    S. R. Mao    H. G. Wang 《Plant Breeding》2007,126(1):9-12
The behaviour of diverse Triticum genotypes in the tissue culture response of mature embryo callus was compared, and factors affecting tissue culture response were studied in this paper. Significant differences were detected in callus induction, embryogenic callus differentiation, plantlet regeneration and culture efficiency when mature embryos of 31 plants of different Triticum species were compared. These were the main wheat cultivars of the Chinese northern winter‐type wheat region and breeding lines (Triticum aestivum L.), durum wheat (Triticum durum Desf.), cultivable emmer wheat (Triticumdicoccum Schuble) and the common wheat progenitors Triticum dicoccoides and Triticum aegilopides. The genotype dependency was particularly high in tissue culture of mature embryos of these Triticum genotypes. The efficiency of induction, differentiation and regeneration of mature embryos callus was high in genotypes selected out. Mature embryo‐derived callus of HB341, TS021, SN2618, T. dicoccum, HB188, and T9817 showed better tissue culture response than the other genotypes. Plantlets can be regenerated from mature embryo‐derived callus of 31 genotypes, saving on growth facility resources and time required for the collection of other explants, and providing a solid basis for the genetic transformation and molecular plant breeding of Triticum plants.  相似文献   

7.
Disorganised embryogenic tissues have been utilised as target tissues for transgene insertion and transgenic plant regeneration in cassava (Manihot esculenta). The production of friable embryogenic callus in fourteen geographically diverse cassava cultivars, from which eleven were established as embryogenic suspension cultures, is reported. Embryogenic tissues were similar in nature in all cultivars tested although there was variation in the time required to generate friable callus and the growth rates of suspension cultures. Regeneration of plants has been achieved from eight cultivars but varied significantly in efficiency, with cv. TMS 60444and Line 2 from Zimbabwe being the most responsive. Tissues from the remaining eight cultivars became arrested at globular and torpedo stages of regeneration indicating that they most likely process an inherent ability to produce plants but require further research to allow this to be realised. Significant numbers of transgenic plants containing transgenes for putative resistance to important viral diseases of cassava in addition to visual marker genes have been regenerated. Transgenic plants from three the cultivars TMS 60444, Bonoua Rouge and M.Col 1505 were recovered after particle bombardment of embryogenic suspension cultures. Correlation's have been made between abnormal leaf morphology and plant vigour with the use of embryogenic suspension cultures for transgene insertion. As an result friable embryogenic callus is now being successfully utilsed as the target tissue for genetic transformation and plant regeneration at ILTAB. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

8.
建立成熟的外植体再生体系,搭建有效的转化平台是实现小麦基因工程改良的重要途径。与未成熟胚相比,利用小麦成熟胚作为外植体不仅取材方便,而且不受环境和季节的限制。因此,建立小麦成熟胚再生体系是小麦组织培养发展的重要趋势。本研究利用"济麦"系列5个品种济南17、济麦19、济麦20、济麦21和济麦22,以适于组培的小麦品种Bobwhite作对照,研究了不同浓度2,4-D、玉米素(ZT)和激动素(KT)对小麦成熟胚的愈伤诱导和分化成苗的影响。结果表明,与对照品种Bobwhite相比,"济麦"系列品种具有更高的成熟胚出愈率,品种间差异显著,呈现出基因型效应。在相同2,4-D诱愈浓度条件下,不同品种间分化和成苗率差异极显著。"济麦"系列品种的成苗率均低于对照品种Bobwhite(13.55%),说明进一步优化愈伤的再生成苗能力是未来建立再生体系的关键。相同品种条件下,2,4-D诱愈浓度对出愈、分化和成苗率影响显著,当2,4-D诱愈浓度4mg/L时,除济麦20以外,其它五个品种均能分化成苗,其中济麦22成苗率约10.33%,接近Bobwhite水平。本研究为建立和完善"济麦"系列品种的成熟胚再生和遗传转化体系奠定了基础。  相似文献   

9.
小麦成熟胚的组织培养   总被引:5,自引:0,他引:5  
【研究目的】为了筛选出小麦(Triticum aestivum L.)成熟胚(MEs)培养和遗传转化受体材料的优良基因型,【方法】以27份优良普通小麦冬性主栽品种或高代育种品系为试材,采用整粒切胚诱愈方法(endosperm-supported method),对小麦成熟胚进行组织培养研究。【结果】结果表明,在成熟胚培养过程中光照、温度、培养基2,4-D浓度、基因型等因素对小麦成熟胚组织培养影响较大;同时小麦成熟胚组织培养存在着较强的基因型依赖性,不同基因型愈伤组织诱导率、分化率、成苗率的差异具有统计学意义(P<0.01)。通过研究从27个小麦基因型中筛选出了3个适合于成熟胚整粒切胚诱愈的基因型,即山农2618、泰山021、河北341,他们的成苗率分别达到了15.38%、25.27%、21.33%。【结论】小麦成熟胚组织培养优良基因型的筛选,为以后小麦的遗传转化和应用基因工程技术进行遗传改良奠定了基础。  相似文献   

10.
Summary Twenty-five inbred lines, including grain and forage types from the USA and China, two hybrids, one Sorghum almum, and one Parasorghum (S. versicolor) were tested for their response to anther culture. Three nutrient media were effective in inducing anther calli from six cultivars (Xin White, TX 403-TSB, DDY Sommer Milo, TX 2779, Brawley, and Spur Federal) and one was effective for plant regeneration for one cultivar, Xin White. Averaged over media, callus induction frequency (number of calli per 100 anthers) was highest in cultivars Xin White and TX 403-TSB (6.7 and 3.9%, respectively). The means of cultivars for media C17-2 and Ms-t-z-2, 4.3 and 3.2%, respectively, were superior to that for medium 85D3-2 (0.1%). Expressed as an average of the six cultivars and three media the mean calli induction frequency was 2.6%; however, differential responses of genotype and medium were noted. Among the 10 regeneration media tested, medium MS-d-4 containing Murashige and Skoog basal components plus 2.0 mg/l indole-3-acetic acid (IAA) and 2.5 mg/l kinetin was the most effective for plant regeneration. Numbers of albino plants and calli developing only roots increased directly with callus-induction time, whereas the frequency of plant regeneration decreased. Regenerated plants had varied numbers of chromosomes in root tip cells: 10, 15, 20, 40, and 60. The 29 regenerated plants that reached maturity, however, were highly fertile and contained only 10 bivalents in pollen mother cells. Normal chromosome number and behavior for the regenerated plants suggest that induced calli originated from cells other than microspores. However, spontaneous chromosome doubling in microspore-derived haploids may occur. The appearance of albinos also implies that haploids may have been produced from anther culture.Joint contribution of the Dept. of Agronomy and USDA-ARS, Kansas Agricultural Experiment Station, Manhattan, KS 66506-5501, USA. Contribution no. 88-566-J.  相似文献   

11.
M. Özgen    M. Türet    S. Özcan  C. Sancak 《Plant Breeding》1996,115(6):455-458
Seven genotypes of winter durum wheat (Triticum durum Desf.) were cultured to establish an efficient method of callus formation and plant regeneration from mature embryo culture, and to compare the responses of immature and mature embryo cultures. Immature embryos were aseptically dissected from seeds and placed, with the scutellum upwards, in dishes containing Murashige and Skoog's (MS) mineral salts and 2mg 2,4- dichlorophenoxyacetic acid (2,4-D) per litre. Calli and regenerated plants were maintained on 2,4-D-free medium. Mature embryos were moved slightly on the imbibed seeds. For callus formation, the seeds with moved embryos were placed, furrow downwards, in dishes containing 8 mg 2,4-D per litre. The developed calli and regenerated plants were maintained on the MS medium. Plants regenerated from both embryo cultures were vernalized and grown to maturity in soil. Variability was observed among the wheat genotypes tested for various culture responses in both explant cultures. Callus induction rate and regeneration capacity of callus were independent of each other. Mature embryos have a low frequency of callus induction but a high regeneration capacity. Considering availability, rapidity and reliability, this form of mature embryo culture can be used as an alternative method for immature embryo culture.  相似文献   

12.
Summary NaCl-tolerant calli were selected from two Japonica and two Indica rice (Oryza sativa L.) cultivars on basal media containing 6,000, 9,000, 12,000 or 15,000 ppm NaCl. Frequency of callus formation decreased with the increase of NaCl in the medium, especially in Indica. About half of the calli of Japonica cultivars selected on NaCl-ammended media survived 20,000 ppm NaCl but none of the Indica callus survived. In Japonica, more plants were regenerated from calli selected on all concentrations of NaCl media than from NaCl-free medium. Concentration of Cl- in callus increased dramatically with increased NaCl content but peroxidase activity decreased.  相似文献   

13.
Summary Genotypic effects on callus induction and plant regeneration in callus, suspension and protoplast culture, and their correlations with both phenotypic and GCA-values for anther culture response, were studied using 21 genotypes of perennial ryegrass. Differences between genotypes accounted for approximately 40% of the total variation for callus induction and initial callus growth, and 59 and 83% of the variation in callus culture for regeneration percentage and percentage of green plants. Effects of genotypes were less pronounced in suspension culture, where suspensions from the same genotype often behaved differently. Some suspension cultures retained their capacity for green plant regeneration for almost two years, repeatedly producing 80–100% green regenerants during this period. Genotypes with high regeneration percentage and a large proportion of green plants from callus culture were also superior in suspension culture for both regeneration performance and longevity. Regeneration percentage and percentage of green plants were uncorrelated, and probably under different genetic control. While capacity for green plant formation from the different genotypes showed no correlation between anther culture and somatic in vitro culture, a positive correlation was observed between the regeneration percentages in somatic in vitro culture and anther culture (r=0.44*–0.85***), suggesting some common genetic control of the two systems.  相似文献   

14.
M. N. Barakat 《Euphytica》1990,48(2):103-110
Summary Callus induction derived from root, hypocotyl and cotyledon explants were investigated for four cultivars of Trifolium alexandrinum L. The Murashige & Skoog (1962) medium containing 2.0 mg/L NAA and 0.5 mg/L BAP was able to induce callus from different explants. A wide range of culture media were tested to determine the morphogenetic potential of different callus types derived from different explant types. Shoot morphogenetic development was observed with the cultivars Sakha 4 and Giza 10. Cell suspension cultures were established from hypocotyl derived callus. Methods for isolation and culture of protoplasts from cotyledons are described.  相似文献   

15.
基因型对陆地棉花药离体培养反应的影响   总被引:3,自引:0,他引:3  
对27种棉花基因型的花药在离体培养中的反应进行了比较,结果表明:所有的基因型在合适的培养条件下均能高频率地形成愈伤组织,但不同基因型间愈伤组织诱导率和诱导量不同;再生植株的花药比原始亲本的花药易诱导出愈伤组织;品种间杂种表现出部分优势,但不明显。本试验中,仅鲁棉6号和Siokra1-3两个品种诱导获得了胚性愈伤组织、胚状体和再生植株,且二者之间也存在着差别。从基因型在棉花花药培养中的效应、外界调控对棉花不同基因型花药培养效果的影响、棉花花药培养与体细胞培养基因型差异的比较等三个方面进行了讨论,提出了棉花花药培养愈伤组织诱导和植株再生是受两套遗传基因控制的遗传性状,并提出了棉花组织培养植株再生的阈值问题  相似文献   

16.
Summary This article reports the culture and plant regeneration of Tripsacum dactyloides. Mature embryos of Tripsacum dactyloides dactyloides were used to obtain embryogenic callus cultures. Currently, 180 normal plants have been regenerated from these cultures. Callus was initiated on MS medium supplemented with dicamba (10 mol or 20 mol) and sucrose (3% or 6%), and plants were regenerated on hormone free MS medium containing 2% sucrose. No significant differences were found in callus initiation frequency or in embryogenic response of cultures on the four combinations of sucrose and dicamba tested. The embryogenic cultures have been maintained for 9 months (12 subcultures) and have retained regeneration capacity. Plants regenerated from tissue culture of maize-by-Tripsacum hybrids could be useful in maize improvement.  相似文献   

17.
A plant regeneration system from rice protoplasts using calli derived from mature embryos was established for the two Brazilian modern rice cultivars IAC-201 and IAC-165. After 30 to 40 days of in vitro culture it was possible to obtain on average 6 million protoplasts per gram of callus. Microscopic selection of embryogenic calli was a key step for protoplast isolation. The production of embryogenic calli increased when L-proline and casein hydrolysate were used in the callus induction medium. The Oc or IR52 nurse cell lines were essential for protoplast division. Different regeneration media were studied and 139 plants were regenerated which set seed. Some of the regenerated plants showed morphological variation such as the presence of awns in spite of the short time of the in vitro culture. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

18.
随着植物抗逆性研究和植物转基因技术的发展,通过异源目的基因转化培育耐盐碱苜蓿品种的研究已引起人们的关注,植物受体高频再生体系的建立是异源转化高效的基础。选取新疆大叶紫花苜蓿种子萌发5~7d无菌苗的子叶、下胚轴及根为外植体,诱导愈伤培养基为MS+2,4-D 0.1~3.0 mg/L(8种不同水平)或MS+2,4-D 2.0 mg/L+ KT 0.01~0.5 mg/L(10种不同水平),诱导芽培养基为MS+6-BA 0.5 mg/L+ NAA 0.05 mg/L,生根培养基为MS。结果表明,外植体在MS+2,4-D 2.0 mg/L+ KT 0.2 mg/L培养基中能够产生状态较好可再分化的愈伤组织,子叶、下胚轴、根的平均出愈率分别为93.1%、100%、100%。愈伤组织在MS+6-BA 0.5 mg/L+ NAA 0.05 mg/L培养基中培养40~80d中均可分化芽,子叶、下胚轴、根的芽平均分化率为50%、78%、50%,将2 cm以上的芽转入MS培养基中诱导生根,14d后,生根的小植株炼苗移入花土中,成活率达90%以上。子叶、下胚轴、根在该体系中均能获得再生植株,根也是一种较好的植株再生材料,以根为外植体进行植株再生的研究报道还较少。  相似文献   

19.
杨跃生 《作物学报》2006,32(8):1204-1208
虽然已经有大豆下胚轴再生植株的很多报道,但再生率总是很低并且不定芽的再生都是在添加细胞分裂素的培养基上取得的。这种培养基使外植体形成大量愈伤组织且阻碍了再生芽的进一步生长。为此本实验将表面灭菌的成熟种子置2种培养基上萌发,一种添加2 mg/L苄基腺嘌呤(BA)而另一种不添加;6 d后将小苗的下胚轴切下作为外植体培养到两种诱导不定芽再生的培养基上,同样一种添加2 mg/L BA而另一种不添加。在9个当地、2个美国和1个巴西共12个供试品系中,最好的培养结果是在含2 mg/L BA的培养基上萌发、长 苗、切取下胚轴并在不含BA的再生培养基诱导不定芽。以此程序,不定芽的再生率和质量都得到了显著提高,其中一个当地品种“广大粒”的再生率达到了72%。  相似文献   

20.
低酚陆地棉直接体细胞胚胎发生和植株再生   总被引:2,自引:0,他引:2  
选用低酚陆地棉无菌苗下胚轴为材料进行全固体组织培养,直接诱导获得了胚性愈伤组织,并进一步分化为再生植株.结果表明,激素是影响棉花直接体细胞胚胎发生的重要因素.MSB培养基中添加2,4-D有利于愈伤组织的形成,却不能直接诱导获得胚性愈伤组织.MSB培养基中添加IBA和BA也不能直接诱导获得胚性愈伤组织.MSB培养基附加适当浓度的IBA和KT能直接诱导出胚性愈伤组织.最适激素组合(1.0 mg/L IBA,0.5 mg/L KT)能使诱导棉花下胚轴产生大量胚性愈伤组织,并且在3个月内就可肉眼观察到不同发育时期的胚.MSB培养基中附加1.0 g/L谷氨酰胺和0.5 g/L天门冬酰胺有利于胚萌发成苗.本研究建立了简便高效的棉花直接体细胞胚胎发生和植株再生培养体系,从胚性愈伤组织诱导到植株再生约需5~6个月时间.  相似文献   

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