以超甜玉米自交系S1和品种粤甜3号为材料研究了影响愈伤组织诱导和胚性形成的几个关键因素。结果表明,2种基因型都是高诱导力的材料,自交系S1能够诱导出高质量的Ⅱ型胚性愈伤,是理想的基因转化材料。2,4-D浓度为2 m g/L时,2种基因型出...以超甜玉米自交系S1和品种粤甜3号为材料研究了影响愈伤组织诱导和胚性形成的几个关键因素。结果表明,2种基因型都是高诱导力的材料,自交系S1能够诱导出高质量的Ⅱ型胚性愈伤,是理想的基因转化材料。2,4-D浓度为2 m g/L时,2种基因型出愈率和胚性愈伤率达到最高,胚性愈伤率分别达到45.88%和35.00%;在培养基中添加9 m g/L的A gNO3极显著地提高了胚性愈伤率,比对照提高了31.25%;添加700 m g/L的脯氨酸,胚性愈伤率显著地提高了12.41%。据此建立了高效稳定的超甜玉米转基因胚性愈伤组织诱导体系。展开更多
Seeds from Tabebuia roseo-alba lose viability very fast. Moreover, the seed germination rate is very low, reaching approximately 40%. This study aimed at the in vitro induction of embryogenic callus. This technology a...Seeds from Tabebuia roseo-alba lose viability very fast. Moreover, the seed germination rate is very low, reaching approximately 40%. This study aimed at the in vitro induction of embryogenic callus. This technology allows subsequent plant regeneration as an alternative for the production of T. roseo-alba seedlings. Seeds were germinated in vitro and after 20 days, cotyledonary leaves, hypocotyls and root segments excised from these seedlings were used as explants. They were inoculated on MS medium supplemented with sucrose （30 g/L）, agar （5.0 g/L） and different auxins. The effect of 2,4-D, picloram and NAA at concentrations 0.0, 0.5, 1.0, 2.0 and 4.0 mg/L was evaluated. For the analysis of callus with embryogenic characteristics, ultra-structural study by scanning electron microscopy and cytochemical test with carmine were performed. The results showed that the culture medium supplemented with 4 mg/L NAA presented induction of callus with embryogenic characteristics in all explants used, with cotyledonary leaves showing the highest percentage （70% of explants with embryogenic characteristics）. The use of 2, 4-D and picloram was efficient for callus formation in different explants, but no embryogenic characteristics were observed. From the ultra-structural analysis of callus with embryogenic characteristics, it was found that cells from different explant sources had isodiametric format. This format is similar to somatic embryos in globular stage. The cytochemical analysis confirmed the presence of pro-embryogenic cells in callus mass. Callus induced from cotyledonary leaves presented 46% positive reaction to carmine acetic.展开更多
Maize is one of the most important cereal crops in Sub-Saharan Africa and an important source of energy for humans. However, the difference in the dedifferentiation frequency of immature embryos among various genotype...Maize is one of the most important cereal crops in Sub-Saharan Africa and an important source of energy for humans. However, the difference in the dedifferentiation frequency of immature embryos among various genotypes indicates that callus induction and genetic transformation is dependent on the genotype. This phenomenon is an impediment in the fundamental process of improving tropical maize germplasm especially through genetic engineering. Here, five tropical maize （Zea mays L.） genotypes, CML 216, CML 144, A 04, E 04 and TL 21, were evaluated for callus induction on MS medium supplemented with the growth regulator dicamba. Embryogenic and non embryogenic callus induction was independent ofgenotype when young immature embryos, 12 days after pollination （DAP） were used for tissue culture in combination with dicamba. The optimal concentration of dicamba for induction ofembryogenic callus in all the genotypes was 3 mg/L, which was also the concentration at which non embryogenic callus formation was lowest. The frequency of embryogenic callus induction ranged from 35% to 79% among the five genotypes and somatic embryos regenerated R0 shoots that produced normal R1 progenies. This regeneration method is expected to facilitate the development of a more efficient genotype independent Agrobacterium- mediated transformation system for tropical inbred lines.展开更多
[Objective] This study was conducted to evaluate the efficiency of somatic embryogenesis and plant regeneration for an upland cotton cultivar Xinluzao33 under the induction of different hormone combinations and thus t...[Objective] This study was conducted to evaluate the efficiency of somatic embryogenesis and plant regeneration for an upland cotton cultivar Xinluzao33 under the induction of different hormone combinations and thus to determine the optimal hormone combination. [Method] Calli of Xinluzao33(Gossypium hirsutum L.) were induced from seedling hypocotyl tissue by a range of DK and BK combinations. Embryogenic calli and embryos were induced on callus-inducing medium(CIM) without any hormones. Callus appearance and quality were compared to determine which medium was the optimal for callus induction. Embryogenesis ratio was calculated to determine which medium was the best for somatic embryogenesis and plant regeneration. [Result] Callus induction rate was 100% in all the 12 hormone combinations.The calli were yellow or kelly, and their texture was loose or soft under low concentrations of DK combinations, green or white, variably compact under high concentrations of DK combinations. The calli induced by BK combinations were kelly or green, covering creamy white substance. The best medium for callus induction was DK6(0.05 mg/L 2, 4-D and 0.10 mg/L KT). Embryogenic calli were successfully induced from all the combinations. The efficiency of embryogenic callus induction,embryogenesis, and plantlet regeneration were significantly different among the 12 combinations. The result showed that the embryogenesis ratio was the highest in BK3 combination(0.50 mg/L IBA and 0.50 mg/L KT), 72.86% of embryogenic calli differentiated into somatic embryos after being cultured on CIM for 80 d, and80.93% of the somatic embryos finally regenerated into plants on SEM(somatic embryo induction medium). [Conclusion] These results indicate that hormone combination BK3(0.50 mg/LIBA and 0.50 mg/L KT) was the best medium for somatic embryogenesis and plant regeneration from Xinluzao33.展开更多
文摘以超甜玉米自交系S1和品种粤甜3号为材料研究了影响愈伤组织诱导和胚性形成的几个关键因素。结果表明,2种基因型都是高诱导力的材料,自交系S1能够诱导出高质量的Ⅱ型胚性愈伤,是理想的基因转化材料。2,4-D浓度为2 m g/L时,2种基因型出愈率和胚性愈伤率达到最高,胚性愈伤率分别达到45.88%和35.00%;在培养基中添加9 m g/L的A gNO3极显著地提高了胚性愈伤率,比对照提高了31.25%;添加700 m g/L的脯氨酸,胚性愈伤率显著地提高了12.41%。据此建立了高效稳定的超甜玉米转基因胚性愈伤组织诱导体系。
文摘Seeds from Tabebuia roseo-alba lose viability very fast. Moreover, the seed germination rate is very low, reaching approximately 40%. This study aimed at the in vitro induction of embryogenic callus. This technology allows subsequent plant regeneration as an alternative for the production of T. roseo-alba seedlings. Seeds were germinated in vitro and after 20 days, cotyledonary leaves, hypocotyls and root segments excised from these seedlings were used as explants. They were inoculated on MS medium supplemented with sucrose （30 g/L）, agar （5.0 g/L） and different auxins. The effect of 2,4-D, picloram and NAA at concentrations 0.0, 0.5, 1.0, 2.0 and 4.0 mg/L was evaluated. For the analysis of callus with embryogenic characteristics, ultra-structural study by scanning electron microscopy and cytochemical test with carmine were performed. The results showed that the culture medium supplemented with 4 mg/L NAA presented induction of callus with embryogenic characteristics in all explants used, with cotyledonary leaves showing the highest percentage （70% of explants with embryogenic characteristics）. The use of 2, 4-D and picloram was efficient for callus formation in different explants, but no embryogenic characteristics were observed. From the ultra-structural analysis of callus with embryogenic characteristics, it was found that cells from different explant sources had isodiametric format. This format is similar to somatic embryos in globular stage. The cytochemical analysis confirmed the presence of pro-embryogenic cells in callus mass. Callus induced from cotyledonary leaves presented 46% positive reaction to carmine acetic.
文摘Maize is one of the most important cereal crops in Sub-Saharan Africa and an important source of energy for humans. However, the difference in the dedifferentiation frequency of immature embryos among various genotypes indicates that callus induction and genetic transformation is dependent on the genotype. This phenomenon is an impediment in the fundamental process of improving tropical maize germplasm especially through genetic engineering. Here, five tropical maize （Zea mays L.） genotypes, CML 216, CML 144, A 04, E 04 and TL 21, were evaluated for callus induction on MS medium supplemented with the growth regulator dicamba. Embryogenic and non embryogenic callus induction was independent ofgenotype when young immature embryos, 12 days after pollination （DAP） were used for tissue culture in combination with dicamba. The optimal concentration of dicamba for induction ofembryogenic callus in all the genotypes was 3 mg/L, which was also the concentration at which non embryogenic callus formation was lowest. The frequency of embryogenic callus induction ranged from 35% to 79% among the five genotypes and somatic embryos regenerated R0 shoots that produced normal R1 progenies. This regeneration method is expected to facilitate the development of a more efficient genotype independent Agrobacterium- mediated transformation system for tropical inbred lines.
基金Supported by the National Transgenic Major Project of China(2009ZX08009-090B)the Biological Germplasm Project of Xinjiang Production&Construction Crops(2012BD046)the Technology Plan of Xinjiang Academy of Agricultural and Reclamation Sciences(81YYD201506)
文摘[Objective] This study was conducted to evaluate the efficiency of somatic embryogenesis and plant regeneration for an upland cotton cultivar Xinluzao33 under the induction of different hormone combinations and thus to determine the optimal hormone combination. [Method] Calli of Xinluzao33(Gossypium hirsutum L.) were induced from seedling hypocotyl tissue by a range of DK and BK combinations. Embryogenic calli and embryos were induced on callus-inducing medium(CIM) without any hormones. Callus appearance and quality were compared to determine which medium was the optimal for callus induction. Embryogenesis ratio was calculated to determine which medium was the best for somatic embryogenesis and plant regeneration. [Result] Callus induction rate was 100% in all the 12 hormone combinations.The calli were yellow or kelly, and their texture was loose or soft under low concentrations of DK combinations, green or white, variably compact under high concentrations of DK combinations. The calli induced by BK combinations were kelly or green, covering creamy white substance. The best medium for callus induction was DK6(0.05 mg/L 2, 4-D and 0.10 mg/L KT). Embryogenic calli were successfully induced from all the combinations. The efficiency of embryogenic callus induction,embryogenesis, and plantlet regeneration were significantly different among the 12 combinations. The result showed that the embryogenesis ratio was the highest in BK3 combination(0.50 mg/L IBA and 0.50 mg/L KT), 72.86% of embryogenic calli differentiated into somatic embryos after being cultured on CIM for 80 d, and80.93% of the somatic embryos finally regenerated into plants on SEM(somatic embryo induction medium). [Conclusion] These results indicate that hormone combination BK3(0.50 mg/LIBA and 0.50 mg/L KT) was the best medium for somatic embryogenesis and plant regeneration from Xinluzao33.
