共查询到18条相似文献，搜索用时 156 毫秒
1.
《分子植物育种》2017,(4)
通过组织培养建立乌菜离体再生体系,可使优良珍贵育种材料得到保存,并能为其快速繁殖提供有效途径。本研究以4种不同基因型乌菜为材料,从外植体类型、基本培养基类型、植物生长调节剂配比、基因型及无菌苗苗龄等方面,对影响乌菜不定芽诱导的因素进行了研究。研究表明,7-1基因型乌菜5 d苗龄的带柄子叶外植体,在以MS为基本培养基(3%蔗糖+0.8%琼脂)添加4 mg/L 6-BA与0.5 mg/Lα-NAA的诱导培养基上进行诱导,不定芽诱导率可达85.56%。本研究对影响乌菜植株再生的多个因素进行了优化,建立了高效离体再生技术体系,为乌菜育种研究及其他生物技术研究提供基础。 相似文献
2.
不结球白菜亮白叶再生体系的优化 总被引：1，自引：1，他引：0
摘 要：本文以不结球白菜品种亮白叶为实验材料,研究不同外植体类型和不同生长调节剂配比等因素对其不定芽分化频率的影响。结果表明,对不结球白菜品种中再生比较困难的品种亮白叶来说,以下胚轴、子叶及子叶柄为外植体的诱导频率存在明显差异。以生长5-6天无菌苗的子叶柄为外植体,在培养基为MS+6-BA（5 mg/L）+NAA（0.1 mg/L）+ AgNO3（4mg/L）的培养条件下,亮白叶分化不定芽的再生频率最高,可达59.5%;这为将来该不结球白菜转化体系的建立奠定了坚实基础。 相似文献
3.
《分子植物育种》2017,(12)
以147花椰菜和庆农65天2个花椰菜品种为试材,研究了影响花椰菜不定芽再生的各个因素,建立花椰菜高效离体再生体系。结果表明:基因型、外植体类型、激素的类型和配比等是影响花椰菜再生的主要因子。品种对再生频率影响较大,庆农65天的外植体再生频率高于147花椰菜;不同外植体再生频率也存在差异,下胚轴的再生频率显著高于子叶。147花椰菜取用7 d龄无菌苗的子叶和下胚轴,培养在MS+6-BA1 mg/L+NAA 0.2 mg/L培养基上培养效果最佳,庆农65天则培养在MS+6-BA 0.5 mg/L+NAA 0.05 mg/L培养基上效果最佳;再生芽在MS培养基,生根率达100%。本研究为花椰菜基因遗传转化构建高效的再生体系奠定基础。 相似文献
4.
利用根癌农杆菌介导法,以白细胞介素-4(interleukin-4,IL-4)基因转化中甘11号甘蓝.本研究对可能影响il-4转化率的外植体类型、外植体的预培养时间、农杆菌的侵染时间以及外植体与农杆菌的共培养时间进行了优化.试验结果表明:il-4基因对带1～2 mm子叶柄的子叶的转化率显著高于下胚轴;带柄子叶在预培养1 d、侵染3 min、共培养1 d时,转化效果最好,转化率达23.3%;下胚轴在预培养1～3 d,侵染3～5 min,共培养1～2 d时,转化效果较好,转化率最高可达13.3%.经PCR检测及PCR-Southern杂交,初步证明目的基因il-4已经转入甘蓝的再生植株.转il-4基因甘蓝的PCR阳性再生植株已经开花并产生了后代. 相似文献
5.
优良再生体系的建立是农杆菌介导植物遗传转化的基础。本研究建立以甘蓝型油菜含部分子叶节的子叶为外植体的再生体系,该再生体系为:切取5日苗龄甘蓝型油菜含部分子叶节的带柄子叶为外植体,置于添加3 mg/L6-BA的MS基本培养基中培养5天,外植体再生出丛生不定芽,不定芽再生率为100%。基于新建立的带节子叶再生体系,通过农杆菌介导,成功地将用p FGC5941载体构建的Bn TFL1基因干扰载体转入甘蓝型油菜中,从播种到得到生根抗性苗,整个转化周期只需要60~70天,较传统甘蓝型油菜以不带节子叶为外植体100~130天的转化周期大大缩短。 相似文献
6.
不同黄瓜基因型子叶再生体系的建立 总被引：1，自引：0，他引：1
以6个不同黄瓜品种和自交系为试材,研究了基因型、激素(6-BA、ABA)对黄瓜离体子叶再生的影响。结果表明:不同基因型黄瓜之间的再生频率差异较大,吉林旱瓜为最佳基因型,在培养基M7(MS+1.5mg/L6-BA+0.5mg/LABA+2mg/LAgNO3)上再生频率最高达97%,平均每外植体再生芽数为5.2;与对照相比,6-BA、ABA和AgNO3组合能促进芽的再生,显著提高再生芽数,其中6-BA是黄瓜子叶再生的必要激素,而ABA是促进子叶高效再生的激素;此外,MS培养基添加0.1mg/L6-BA能显著促进再生不定芽的伸长。 相似文献
7.
花生不定芽分化及植株再生的研究 总被引：2，自引：2，他引：0
为进一步探索和完善高频植株再生体系并扩大基因型的研究,以8个花生品种为材料,对幼叶不同部位（叶片顶端、中间切段、叶片基部）的切段、子叶、上胚轴和下胚轴外植体,接种到添加1 mg/L NAA和6 mg/L BAP的MSB5培养基上,10天后将形成的愈伤组织转移至添加10 mg/L BAP或添加3 mg/L BAP和1 mg/L ABA的再分化培养基上进行培养,诱导不定芽分化。结果表明,6天叶龄叶片中间切段不定芽分化频率达到了91.4%,上胚轴26.7%,下胚轴12.5%和子叶0%;花生品种8823幼叶外植体获得了的91.4%不定芽分化率,生根植株经驯化后移至砂土中,正常开花结果。因此,幼叶外植体分化不定芽频率明显高于子叶、上胚轴和下胚轴;6天叶龄的叶片中间切段为最佳;不同基因型不定芽分化率存在明显差异。 相似文献
8.
本研究建立了以甘蓝型油菜下胚轴为外植体的一步不定芽再生培养和遗传转化体系。首先,以甘蓝型油菜中双11号含部分子叶节的下胚轴为外植体,从6-BA和NAA配比、Ag NO3以及无菌苗苗龄等方面对影响油菜组织培养的因素进行了研究,建立了甘蓝型油菜快速高频一步不定芽再生培养技术体系。该技术体系为,切取5 d苗龄含部分子叶节的下胚轴置于添加4 mg/L 6-BA的MS基本培养基中培养,5 d再生出不定芽,再生频率为100%。在此基础上,进行了甘蓝型油菜的遗传转化,成功地将用p FGC5941双元载体构建的Bn TFL1基因干扰载体转入中双11号中。整个转化过程中,芽的诱导生成只需5 d,接着完成抗性芽的PPT筛选需要30 d,整个转化周期从播种到得到生根抗性苗仅需大约70 d,而传统转化方法抗性芽的诱导筛选需要90 d,整个转化周期需要130 d左右,大大缩短了转化周期,简化了转化过程,提高了转化效率。 相似文献
9.
影响小白菜子叶再生的因素 总被引：3，自引：1，他引：2
本研究以小白菜“杭州油冬儿”品种为试材,研究了不同激素组合、子叶不同部位、不同接种方式对不定芽再生的影响,并且还筛选了种子表面消毒的最佳方法和不定芽生根的最佳培养基。研究结果表明,白菜种子表面消毒的最佳方法为用70%的乙醇浸泡30s,再用20%的次氯酸钠消毒20min或用0.1%升汞消毒10min;以完整的带柄子叶为最佳外植体,接种时将子叶背面朝上放置比正面朝上放置更有利于不定芽的分化;子叶再生的最佳培养基为MN＋BA2mg/L＋NAA0.5mg/L,不定芽生根的最佳培养基为1/2MS＋IAA0.2mg/L。此研究为小白菜再生体系的建立奠定了基础。 相似文献
10.
番茄组培再生体系优化研究 总被引：9，自引：3，他引：6
对番茄再生体系中无菌苗的苗龄、不定芽诱导培养基和生根培养基的激素配比进行了优化。试验以‘浙杂905’、‘圣亚’和‘富丹’为材料。用番茄的茎段和子叶作为外植体,对外植体的出愈率、诱导率和不定芽生根情况进行差异比较。主要结果如下：选苗龄为4天的番茄幼苗。番茄子叶为外植体的最佳再生培养基配比为MSB5+3%蔗糖+0.6%琼脂+BA 2.0 mg/L+NAA 0.1 mg/L（pH5.8）,‘浙杂905’、‘圣亚’和‘富丹’子叶的出愈率分别为97.00%、91.00%、86.00%,诱导率分别为64.67%、59%、53%。以番茄茎段为外植体的最佳再生培养基配比为MSB5+3%蔗糖+0.6%琼脂+BA 2.0 mg/L+NAA 0.2 mg/L（pH5.8）,‘浙杂905’、‘圣亚’和‘富丹’茎段的出愈率分别为81.67%、76.00%、82.67%,诱导率分别为41.00%、48.67%、39.67%。不定芽生根培养基为：1/2MS+NAA 0.1 mg/L +IBA 0.5 mg/L。通过试验对比得出,番茄再生体系中选用无菌苗的最佳苗龄、不定芽诱导培养基和生根培养基的配比,子叶的再生能力要显著高于茎段。 相似文献
11.
Vipasha Verma Kamlesh Kanwar Mahak Tufchi Monika Kashyap 《Journal of Crop Science and Biotechnology》2014,17(1):1-10
Three different regeneration systems, viz. regeneration through callus cultures using embryonic explant, direct regeneration using shoot bud/nodal segments as explant and regeneration through cell suspension culture using cotyledonary explant (for the induction of transgenic callus for suspension culture) were evaluated to see their effect on transfer of Cry1A(b) gene to Punica granatum L. cv. Kandhari Kabuli through Agrobacterium mediated transformation. Pre-conditioning and co-cultivation durations had a marked effect on transformation frequency of different explants. Out of different explants used (embryo, shoot bud, and cotyledon) for different regeneration systems cotyledonary explant showed highest putative transformation frequency (13.54%) inducing callus on selective medium for carrying out cell suspension culture to regenerate transgenic shoots. Despite of the highest transformation frequency obtained from the cotyledon explant, the plating efficiency of the transgenic cells generated through the transgenic callus (callus formed from the cotyledonary explant) during cell suspension culture was found to be very low (0.7%). Thus the plating efficiency has also played worth mentioning role in the regeneration of transformants following cell suspension culture. Among the three regeneration systems, regeneration through callus cultures using embryonic explant was found to be best for regeneration of transformants. The highest per cent regeneration of 23.33 was obtained from the putative transgenic embrogenic calli. Successful genetic transformation in the transformed plantlets was confirmed by PCR analysis. The transformation system thus developed is valuable and may be used to produce insect resistant trees. 相似文献
12.
Medium, Explant and Genotype Factors Influencing Shoot Regeneration in Oilseed Brassica spp. 总被引：9，自引：0，他引：9
G. X. Tang W. J. Zhou H. Z. Li B. Z. Mao Z. H. He K. Yoneyama 《Journal of Agronomy and Crop Science》2003,189(5):351-358
The effects of culture media, explants and genotypes on shoot regeneration in oilseed Brassica species were examined in this study. The maximum shoot regeneration frequency was obtained in Murashige and Skoog medium supplemented with 3 mg l?1 6‐benzylaminopurine and 0.15 mg l?1 1‐naphthaleneacetic acid. The addition of 2.5 mg l?1 AgNO3 was very beneficial to shoot regeneration in B. napus and Ag2S2O3 (10 mg l?1) was even superior to AgNO3 (2.5 mg l?1). Explant age, explant type and carbon source also significantly affected shoot regeneration. Four‐day‐old seedlings of cotyledonary explants showed the maximum shoot regeneration frequency and number of shoots per explant. Of the four explants – peduncles, hypocotyls, cotyledons and leaf petioles – cotyledons produced the highest shoot regeneration frequency (56.67 %). Four carbon sources – glucose, maltose, starch and sucrose – were compared for their respective effects on shoot regeneration from cotyledonary explants. Sucrose appeared to be the best carbon source for shoot regeneration with the highest shoot regeneration frequency (76.00 %). Considerable variation in shoot regeneration from cotyledonary explants was observed both between and within Brassica species. The shoot regeneration frequency ranged from 10.00 % for cv. R5 (B. rapa) to 83.61 % for cv. N1 (B. napus). Two B. napus, one B. carinata and one B. juncea cultivars exhibited shoot regeneration frequency higher than 70 %. In terms of the number of shoots produced per explant, B. rapa showed the highest variation, ranging from 5.64 for cv. R3 to 1.33 for cv. R5. Normal plantlets were regenerated from all induced shoots and developed normally. The regenerated plants were fertile and identical with the source plants. 相似文献
13.
优化通过器官直接再生方式的黄瓜离体再生体系,为遗传转化奠定基础。以‘长春密刺’黄瓜的子叶节为外植体,探讨在黄瓜再生过程中,适宜的无菌苗获得培养基、适宜的外植体类型、无菌苗的适宜苗态、不定芽诱导培养基和芽伸长培养基中适宜的激素组合与比例。结果表明,最适的无菌苗获得培养基为1/2MS+30 g/L蔗糖+7 g/L琼脂(pH=5.8);不同外植体的再生率为子叶节>下胚轴>子叶;子叶完全出壳但未展平的无菌苗比其他苗态的再生率高,子叶展平后,再生率迅速下降;当6-BA（6-苄基氨基嘌呤）和ABA（脱落酸）浓度一定时,最适的AgNO3为2 mg/L;当AgNO3浓度一定时,6-BA和ABA浓度的最佳组合为1.5 mg/L 6-BA+0.5 mg/L ABA;不定芽诱导的最适培养基为MS+1.5 mg/L 6-BA+0.5 mg/L ABA+2 mg/L AgNO3,出芽率为90%,每外植体出芽数为3.5;芽伸长培养基中加入0.10 mg/L 6-BA,能够促进再生芽的伸长。本研究成功优化了黄瓜的再生体系,得到了健壮的黄瓜成株。 相似文献
14.
以黄瓜（Cucumis sativus L.）子叶节为外植体，研究了品种、6-苄氨基嘌呤、外植体大小及硝酸银对黄瓜直接不定芽诱导的影响。结果表明：不同黄瓜品种直接不定芽数量诱导存在显著差异，但长度诱导无显著差异；6-苄氨基嘌呤对直接不定芽数量和长度的诱导作用显著，其中4.0mg/L为数量诱导适宜浓度，其出芽率和每外植体出芽数达到最高，分别为100.0%和11.1，0.5mg/L为长度诱导适宜浓度，其平均芽长12.0mm；外植体的大小对直接不定芽数量和长度均具显著影响，随着子叶的增大诱导直接不定芽的数量和长度增加，其中单片完整子叶为最适宜大小，其出芽率、每外植体出芽数和平均芽长分别达到100%、6.7和17.1mm；硝酸银也对直接不定芽数量诱导影响显著，其中2.0mg/L为适宜浓度，出芽率和每外植体出芽数分别达到97.2%和4.2，但对芽长有抑制作用。 相似文献
15.
为建立高效的组织培养体系，以甘蓝型油菜杂交品种恢复系627R、621R和616R材料田间种植植株的侧芽、花托和无菌种子实生苗下胚轴为外植体，探索不同苗龄、预培养时间、预培养基、愈伤分化培养基、诱导出芽培养基以及成苗壮苗培养基中激素配比对芽再生、成苗植株生长势的影响。结果表明：无菌苗快速繁殖体系中，发芽6 天的无菌苗下胚轴或者子叶在预培养（MS+ 2.0 mg/L 2,4-D+ 1.0 mg/L 6-BA+ 30 g/L 蔗糖+ 8 g/L 琼脂，pH 5.85）3 天后转移到分化培养基MS+ 3 mg/L 6-BA+ 1.0 mg/L NAA+5 mg/L AgNO3+ 30 g/L 蔗糖+8 g/L 琼脂(pH 5.85)，或者MS+ 3 mg/L 6-BA+ 1.0 mg/L IAA+ 5 mg/LAgNO3+ 30 g/L 蔗糖+8 g/L 琼脂(pH 5.85)生长，可以获得较高的芽再生频率，对于田间生长到抽薹开花期植株取样的外植体，腋芽的出芽频率高于花托培养的出芽频率，但是这2 类外植体在分化培养基（MS+ 10 mg/L 6-BA+ 1 mg/L NAA+ 30 g/L 蔗糖+8 g/L 琼脂，pH 5.85）生长30 天后都有成苗的潜力，上
述外植体经过组织培养出芽后转移到添加矮壮素的培养基（MS+15 mg/L CCC+15~20 g/L 蔗糖+8 g/L 琼脂，pH 5.85）上继续生长30 天，能够得到根系发达、生长势强的植株。甘蓝型油菜优良恢复系建立的组织培养体系能够快速获得生长势优的油菜植株，加速油菜良种的繁殖与评价。 相似文献
16.
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
17.
Arpita Moharana Aradhana Das Enketeswara Subudhi Soumendra K. Naik Durga P. Barik 《Journal of Crop Science and Biotechnology》2017,20(5):405-416
An efficient and reproducible protocol for in vitro plant regeneration was developed for Lawsonia inermis L. using cotyledonary node explant derived from axenic seedlings. Highest shoot proliferation frequency (ca 96.6%) was achieved on Murashige and Skoog’s, 1962 (MS) basal medium supplemented with 8.88 μM 6-Benzyladenine (BA) + 2.68 μM Napthalene acetic acid (NAA). Up-scaling of shoots was carried out using in vitro nodes on MS medium supplemented with 4.44 μM BA. So overall, an average of 238 shoots was produced at 75 days. Of the four different forms of cotyledonary node explants evaluated, highest shoot multiplication was observed in cotyledonary node explant with two whole cotyledons. In vitro regenerated shoots were best rooted (ca 34.3 roots / shoot) on ½ MS medium devoid of any growth regulator. The plantlets were successfully acclimated in sand:soil:: 1:1and established in the garden soil. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) analysis revealed a homogeneous amplification profile for all micropropagated plants validating the genetic fidelity of the in vitro-regenerated plants and supporting the regeneration protocol for economic commercial exploitation. 相似文献
18.
Protocols of plant regeneration have been developed for Brassica carinata for creating somaclonal variation for plant type and adaptability, so that this species can fit into cropping systems in Indian agriculture. The response of cotyledonary and stem explants was assessed for callus induction and shoot regeneration on MS and B5 basal media containing different combinations of auxin and cytokinin concentrations. MS medium supplemented with BA and NAA favoured callus induction. Supplementing MS with combinations of BA and IAA, as also with BA alone, regenerated shoots from the ex pi ants with a high frequency. The frequency of shoot regeneration and the mean number of shoots per explant were higher in cotyledons than in stem explants on identical growth regulator combinations. On B5 medium, supplemented with BA (2 mg/l) and IBA (0.4 mg/l), compact callus was produced which regenerated shoots on transfer to medium containing BA (0.8 mg/l). Genotypic differences among carinata accessions for regeneration were also observed. 相似文献