An efficient in vitro protocol for multiple shoot induction in mulberry, Morus alba L variety V1 (original) (raw)
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Scientia Horticulturae, 1999
A high frequency of sprouting (80.0%) and shoot differentiation was observed in the primary cultures of nodal explants of Morus indica L. cultivar M-5 on MS medium supplemented with 2,4-D (0.3 mg/l). In vitro proliferated shoots were multiplied rapidly by culture of shoot tips on MS medium with BAP (0.5 and 1.0 mg/l) which produced the greatest multiple shoot formation. Multiplication was also achieved by culture of shoot tips on MS medium with BAP (4.0 mg/l) and GA 3 (0.05 mg/l) which facilitated the elongation of shoots followed by sprouting of axillary buds of in vitro grown shoots. A high frequency of rooting (86.7%) with development of healthy roots was observed from shoots cultured on medium with 2,4-D (1.0 mg/l). Plants with well developed roots were transferred to soil with a survival frequency of 80%.
In Vitro propagation of mulberry (Morus indica) var. S-1635 from nodal explants
2016
In the present study, nodal explants from mature trees were used for axillary shoot proliferation of Morus indica var S-1635. Nodal explants (1.5cm long) were cultured in MS medium supplemented with different levels of N 6-benzyladenine (BA: 1, 2, 4, 8, 10 or 12µM) and 1-nephthaleneacetic acid (NAA: 1, 2, 3 or 4µM for each). The rate of bud break was 100% at 10µM BA in combination with either concentration of NAA (2 or 3µM) after 6 days of culture. Generally, the length of axillary shoots was significantly improved by increasing the level of BA with auxin. Most of the nodal explants (65.2%) developed surface calluses on the aerial portions of explants at comparatively lower BA levels (2-4µM). The elongating axillary shoots were cut to prepare further nodal explants for multiple shoot induction. MS medium supplemented with BA (8.8µM) and 2µM indole-3-butyric acid (IBA) was quite effective for 65% shoot induction with an average of 4.7 numbers of shoots and 8.4mm shoot length, after a...
Journal of Experimental Biology and Agricultural Sciences, 2018
Morus multicaulis cv. Goshoerami is the leading mulberry variety for silkworm rearing under temperate climatic conditions of Jammu and Kashmir, India. However, the propagation of this popular mulberry variety has always remained a point of contention due to its poor rooting response through stem cuttings. It normally takes 4 to 5 years for raising the saplings of this variety through conventional root grafting techniques. Therefore, for quick propagation of this poor rooting popular mulberry variety, a one step in vitro protocol was developed by culturing nodal explants from 2 year old plants on Murashige & Skoog (MS) media supplemented with individual as well as combination of phytohormones. The maximum shoot bud proliferation (6.3± 0.71 in cm) and rooting (14.7± 0.53 in cm) was observed when nodal explants were cultured on the combinational media of BAP (1 mg/L) and IBA (1 mg/L) after 14 days of culture. These in vitro raised plantlets were hardened by using the sterile soil and vermiculite in 2:1 ratio. Only 25 days were required for the micro propagation and hardening of raised plantlets of Goshoerami through this single step protocol. The hardened plantlets were successfully established in the field with 83% survival rate. The developed one step protocol can be used efficiently for the mass propagation of this elite mulberry variety throughout the year with in short span of 25 days.
A high frequency of sprouting (80%) from nodal-and (70%) from shoot tip explants and shoot differentiation was observed in the primary cultures of Morus alba L. on MS medium supplemented with BAP and Kn. In vitro proliferated shoots were multiplied rapidly by culture of shoot tips and nodal explants on MS with BAP (2 mg/l) and NAA (0.2 mg/l) as supplements. This combination proved best for multiple shoot formation. Multiplication was also achieved by culture of both the kinds of explants on MS fortified with BAP (2 mg/l) + NAA (0.2 mg/l) + aspargine (25 mg/l) + glutamine (1 mg/l). This medium facilitated the elongation of shoots and sprouting of axillary buds of in vitro grown shoots. About 80% rooting was obtained from shoots cultured on the MS supplemented with NAA (1.0 mg/l). Plants with well developed roots were transferred to soil with survival frequency of 70%.
Application of Tissue Culture Techniques for Propagation and Crop Improvement in Mulberry (Morus spp
Mulberry (Morus spp.) is an economically important tree grown widely in China, India and several other countries in Asia. The major economic product of mulberry is its leaf which is used for feeding the monophagous silkworm Bombyx mori L. Mulberry leaf is also used as fodder for livestock. Mulberry fruit is good for human consumption. Although mulberry is amenable to both sexual and asexual methods of reproduction, due to the high heterozygosity of the parental lines and long juvenile periods, propagation is mainly through stem cuttings or bud grafting. However, success of propagation through stem cuttings is greatly dependent on the genotype, environment, and age of the planting materials. Besides, most of the temperate species are hard to root from the stem cuttings. Micropropagation is seen as a cost effective method for propagation of these species, though a number of factors affect the success. In this article, we summarize these factors along with the causes and remedies for them. Other applications of tissue culture such as germplasm conservation, screening for stress tolerance, triploid developments, genetic transformation, and their impacts on the sericulture industry have also been detailed and discussed.
2015
Mulberry (Morus spp.) is a crop of economic importance in the sericulture industry. In present investigation, simple protocol for shoot induction via micropropagation of Morus indica in different types of tissue culture media was developed. All four media were tested with different growth hormones for multiplication stage. MS was found to be the best medium for shoot induction, high frequency of induced shoots were obtained on MS+ BAP (3.0 mg/l). Multiple shoots were also achieved from in-vitro raised shoots on all the four different medium supplemented with BAP (1.0-1.5 mg/l), NAA (0.25-0.5 mg/l) and GA 3 (0.5 mg/l). The highest shoot length (2.12 ± 1.01) with maximum no. of leaves were obtained in MS medium, followed by shoot length (2.0±1.37) in WPM media.
American Journal of Plant Sciences, 2014
Shoot meristems used for the study were exercised from the in vitro regenerated shoots cultured on MS medium supplemented with 0.5 mg/L of BAP for multiplication. The sensitivity of the in vitro regenerated was studied using shoot meristems of 0.5 cm. Shoot meristems were cultured on medium containing 10-100 mg/l kanamycin to determine the concentration that was lethal for multiple shoot induction and root induction. The response of shoot multiplication decreased (66.2%-6.2%) as the concentration of kanamycin increased (10.0-70.0 mg/L) with complete inhibition of shoot proliferation at 100 mg/L kanamycin. The rooting phase was very sensitive to kanamycin compared to shoot multiplication. The percentage of shoots that rooted decreased (53.8%-4.8%) with increase in the concentration of kanamycin (10.0-70.0 mg/l) on IBA and 2,4-D supplemented medium. For transformation studies, the shoot tips that were infected with Agrobacterium strain were placed on selection medium containing MS medium with 0.5 mg/L BAP and 100 mg/L kanamycin and scored for the putative transformed shoots. An average of 62.2% of shoot tips developed shoot buds from the base and the shoots reached a length of 0.5-1.0 cm at the end of 30 days of culture on the selective medium in comparison to control which showed no response. An average of 66.7% of the regenerated plants showed GUS expression on selection medium where 43.2% and 65% of GUS expression was recorded in the leaves and callus. Leaves and callus induced from the controls did not show GUS activity. Stable integration of nptII gene with the genomic DNA from these transformed plants was confirmed through PCR analysis. Our result presents an efficient regeneration system using in vitro derived shoot meristems for Agrobacterium mediated gene transfer.