Cherry regeneration via somatic embryogenesis is a powerful tool to breeding. In this way, the embryogenic capacity of Prunus incisa specie has been tested from leaves under different interactions of picloram concentr...Cherry regeneration via somatic embryogenesis is a powerful tool to breeding. In this way, the embryogenic capacity of Prunus incisa specie has been tested from leaves under different interactions of picloram concentrations and darkness exposures. Induction culture was achieved on MS medium supplemented with picloram concentrations at 0.5, 1 and 1.5 mg.L1 and submitted to 10, 20, 30 and 40 days of darkness. The best rate of embryogenic leaves was obtained with the interaction of 30 days darkness exposure＊ 1 mg.L^-1 picloram. According to their age, leaves were differently reacted to somatic embryogenesis; indeed, the 2nd expanded leaf from the apex was the most embryogenic one. Concerning the effect of additional auxin to picloram （1 mg·L^-1）, IAA at 0.1 mg·L^-1 and IBA at 0.1 mg·L^-1 gave significantly higher induction rates than all other concentrations, but regenerating somatic embryos showed some teratological abnormalities probably due to seconda;y embryogenesis. At the opposite, NAA at 0.5 mg·L^-1 didn＇t improve embryogenic rate but affected positively embryo development. Furthermore, embryogenesis preferentially took place on the basal part of leaf. Satisfactory rates of somatic embryogenesis are obtained but further improvement remains possible.展开更多
The establishment of high frequency regeneration system is a foundation for Agrobacterium mediated genetic transformation. In this work, several important factors influencing the efficiency of regeneration of pla...The establishment of high frequency regeneration system is a foundation for Agrobacterium mediated genetic transformation. In this work, several important factors influencing the efficiency of regeneration of plants, such as concentration of plant growth regulators, leaf explant orientation, leaf growth sequence and leaf segment, were studied. The results indicated that the differentiation rate of adventitious shoots was 90% on basal MS medium only supplemented with 1 5?mg·L -1 BA (6 benzyladenine) and reached the highest level(95%) when 1 0?mg·L -1 BA and 0 3?mg·L -1 NAA (naphthaleneacetic acid) were added to MS medium. 90% of differentiation rate of adventitious roots were obtained when 0 3?mg·L -1 NAA was only added to MS medium. It was also found that more adventitious shoots were regenerated from the lower segment of leaf (with petiole) than the other segments, the number of adventitious shoots decreased from top to base of leaf growth sequence and the percentage of adventitious shoot induction with adaxial side downward was higher than that with adaxial side upward.展开更多
The effects of Timentin and cefotaxime （Cef） on shoot regeneration of the London plane tree （Platanus acerifolia Willd.） and their use for the suppression ofAgrobacterium tumefaciens in Agrobacterium-mediated gene...The effects of Timentin and cefotaxime （Cef） on shoot regeneration of the London plane tree （Platanus acerifolia Willd.） and their use for the suppression ofAgrobacterium tumefaciens in Agrobacterium-mediated genetic transformation were compared. Shoot regeneration was significantly reduced on the media with Cef at concentrations from 100 to 500 mg·L^-1. Timentin showed negative effect on plant regeneration at concentrations of 100 and 500 mg·L^-1; however, 300 mg·L^-1 Timentin was shown to facilitate shoot regeneration significantly and the regeneration frequency increased from 64% （control） to 88%. Effective suppression of A. tumefaciens could be obtained with 500 mg·L^-1 Cef, but plant regeneration was completely inhibited at this level. The A. tumefaciens on infected P. acerifolia leaf tissues was visually undetectable after three subcultures on a medium with 300 mg·L^-1 Timentin. Considering the effect of Cef and Timentin on plant regeneration and suppression of Agrobacteria, Timentin at 300 mg·L^-1 is the preferred application in .4. tumefaciens-mediated transformation ofP acerifolia.展开更多
The effect of different concentrations of 6-benzylaminopurine （BA） with or without 0.2 mg/L NAA on in vitro regeneration of sugarcane （Saccharum spp.） cultivars SP726180, B47419, M1176/77 and M2119/88 were evaluat...The effect of different concentrations of 6-benzylaminopurine （BA） with or without 0.2 mg/L NAA on in vitro regeneration of sugarcane （Saccharum spp.） cultivars SP726180, B47419, M1176/77 and M2119/88 were evaluated. Leaf base explants were cultured on Murashige and Skoog （MS） basal medium supplemented with 3.0 mg/L 2,4-dichlorophenoxyacetic acid （2,4-D） for 4 weeks. Thereafter, induction of somatic embryogenesis was observed following the transfer of resulting calli to 2,4-D-free medium for another 4 weeks. Regeneration was achieved by transfer of the embryogenic calli to regeneration media fortified with different concentrations of BA ＋ tt-naphthylacetic acid （NAA）. The number, length and vigor of shoots produced in all the genotypes were highest on media supplemented with 1.0 and 1.5 mg/L BA with and without 0.2 mg/L NAA. Among the genotypes tested, B47419 and M1176/77 recorded highest number of shoots, while maximum shoot length and crop vigor was obtained with M1176/77. Induction of callus with 3.0 mg/L 2,4-D and its subsequent incubation on 2,4-D-free media, followed by regeneration on media supplemented with 1.0 or 1.5 mg/L BA with 0.2 mg/L NAA was found to be efficient for in vitro regeneration of the sugarcane genotypes used in this study. This protocol could be applied for micropropagation of other elite genotypes.展开更多
文摘Cherry regeneration via somatic embryogenesis is a powerful tool to breeding. In this way, the embryogenic capacity of Prunus incisa specie has been tested from leaves under different interactions of picloram concentrations and darkness exposures. Induction culture was achieved on MS medium supplemented with picloram concentrations at 0.5, 1 and 1.5 mg.L1 and submitted to 10, 20, 30 and 40 days of darkness. The best rate of embryogenic leaves was obtained with the interaction of 30 days darkness exposure＊ 1 mg.L^-1 picloram. According to their age, leaves were differently reacted to somatic embryogenesis; indeed, the 2nd expanded leaf from the apex was the most embryogenic one. Concerning the effect of additional auxin to picloram （1 mg·L^-1）, IAA at 0.1 mg·L^-1 and IBA at 0.1 mg·L^-1 gave significantly higher induction rates than all other concentrations, but regenerating somatic embryos showed some teratological abnormalities probably due to seconda;y embryogenesis. At the opposite, NAA at 0.5 mg·L^-1 didn＇t improve embryogenic rate but affected positively embryo development. Furthermore, embryogenesis preferentially took place on the basal part of leaf. Satisfactory rates of somatic embryogenesis are obtained but further improvement remains possible.
文摘The establishment of high frequency regeneration system is a foundation for Agrobacterium mediated genetic transformation. In this work, several important factors influencing the efficiency of regeneration of plants, such as concentration of plant growth regulators, leaf explant orientation, leaf growth sequence and leaf segment, were studied. The results indicated that the differentiation rate of adventitious shoots was 90% on basal MS medium only supplemented with 1 5?mg·L -1 BA (6 benzyladenine) and reached the highest level(95%) when 1 0?mg·L -1 BA and 0 3?mg·L -1 NAA (naphthaleneacetic acid) were added to MS medium. 90% of differentiation rate of adventitious roots were obtained when 0 3?mg·L -1 NAA was only added to MS medium. It was also found that more adventitious shoots were regenerated from the lower segment of leaf (with petiole) than the other segments, the number of adventitious shoots decreased from top to base of leaf growth sequence and the percentage of adventitious shoot induction with adaxial side downward was higher than that with adaxial side upward.
基金This research is partially supported by the National Natural Science Foundation of China （Grant No. 30371015） the Bureau of Science and Technology of Wuhan. The kindness of doctor Lin Yong-jun in providing Agrobacterium tumefaciens strains EHA 105, harboring the binary vector pCAMBI2301 for this experiment is greatly appreciated. We thank all the colleagues in our laboratory, especially doctor Gao Li-ping, for constructive discussion and technical support.
文摘The effects of Timentin and cefotaxime （Cef） on shoot regeneration of the London plane tree （Platanus acerifolia Willd.） and their use for the suppression ofAgrobacterium tumefaciens in Agrobacterium-mediated genetic transformation were compared. Shoot regeneration was significantly reduced on the media with Cef at concentrations from 100 to 500 mg·L^-1. Timentin showed negative effect on plant regeneration at concentrations of 100 and 500 mg·L^-1; however, 300 mg·L^-1 Timentin was shown to facilitate shoot regeneration significantly and the regeneration frequency increased from 64% （control） to 88%. Effective suppression of A. tumefaciens could be obtained with 500 mg·L^-1 Cef, but plant regeneration was completely inhibited at this level. The A. tumefaciens on infected P. acerifolia leaf tissues was visually undetectable after three subcultures on a medium with 300 mg·L^-1 Timentin. Considering the effect of Cef and Timentin on plant regeneration and suppression of Agrobacteria, Timentin at 300 mg·L^-1 is the preferred application in .4. tumefaciens-mediated transformation ofP acerifolia.
文摘The effect of different concentrations of 6-benzylaminopurine （BA） with or without 0.2 mg/L NAA on in vitro regeneration of sugarcane （Saccharum spp.） cultivars SP726180, B47419, M1176/77 and M2119/88 were evaluated. Leaf base explants were cultured on Murashige and Skoog （MS） basal medium supplemented with 3.0 mg/L 2,4-dichlorophenoxyacetic acid （2,4-D） for 4 weeks. Thereafter, induction of somatic embryogenesis was observed following the transfer of resulting calli to 2,4-D-free medium for another 4 weeks. Regeneration was achieved by transfer of the embryogenic calli to regeneration media fortified with different concentrations of BA ＋ tt-naphthylacetic acid （NAA）. The number, length and vigor of shoots produced in all the genotypes were highest on media supplemented with 1.0 and 1.5 mg/L BA with and without 0.2 mg/L NAA. Among the genotypes tested, B47419 and M1176/77 recorded highest number of shoots, while maximum shoot length and crop vigor was obtained with M1176/77. Induction of callus with 3.0 mg/L 2,4-D and its subsequent incubation on 2,4-D-free media, followed by regeneration on media supplemented with 1.0 or 1.5 mg/L BA with 0.2 mg/L NAA was found to be efficient for in vitro regeneration of the sugarcane genotypes used in this study. This protocol could be applied for micropropagation of other elite genotypes.
