Abstract
BackgroundAloe vera (L.) Burm.f is an important industrial crop, which has enormous application in pharmaceutical, cosmetic and food industries. Thereby, the demand for quality planting material of A. vera is increasing worldwide. Micropropagation is the widely accepted practical application of plant biotechnology that has gained the status of a multibillion-dollar industry throughout the world and this techniques can be used to meet the industrial demand of A. vera. Present studies aim to develop a proficient methods of high-frequency true-to-type plantlet regeneration without intermediate callus phase for A. vera.ResultsNodal portion of rhizomatous stem of A. vera were cultured on Murashige and Skoog (MS) medium (Physiol. Plant. 15:473 – 497, 1962) supplemented with various cytokinin and A. vera leaf gel (AvG) as organic supplement. Number of proliferated shoots per explant was increased along with the regeneration cycles and on MS medium supplemented with 2.5 mg/L 6-benzylaminopurine and 10.0% (v/v) AvG, only 17.8 ± 0.35 shoots per explant were induced on 1st regeneration cycle whereas on 3rd regeneration cycle these number increase to 38.5 ± 0.44 shoots per explant on the same medium composition. AvG have an encouraging role to increase the proliferation rate and on 3rd regeneration cycle 27.6 ± 0.53 shoot per explant induced on 2.5 mg/L BAP, but these number increase to 38.5 ± 0.44 shoots per explant when 10.0% (v/v) AvG was added along with 2.5 mg/L BAP. After transfer of individual excised shoots to a one-third strength MS medium containing 20.0% (v/v) AvG, all the shoots formed whole plantlets with maximum number (9.6 ± 0.29) of roots per shoot. 95.0% of the regenerated plantlets survived on poly-green house. Normal flower appeared in 84.2% field growing micropropagated plants after 18 to 20 months of field transfer. Further, clonal fidelity of the two years old micropropagated plants was established by studying mitotic and meiotic chromosomal behavior and also considered the chromosome number and structural organization. There were no alterations in chromosome phenotypes, somatic haploid (pollen mitosis) and diploid chromosome count (n = 7; 2n = 14), or meiotic behavior. Randomly amplified polymorphic DNA analyses revealed there were no somaclonal variations among these regenerants.ConclusionsThese results confirm the very reliable method for large scale production of true-to-type plantlets of A. vera, which can be used for commercial purpose.Electronic supplementary materialThe online version of this article (doi:10.1186/1999-3110-54-46) contains supplementary material, which is available to authorized users.
Highlights
Aloe vera (L.) Burm.f is an important industrial crop, which has enormous application in pharmaceutical, cosmetic and food industries
Initiation of shoot buds were observed in naked eye within 26-35 d of implantation depending on the types and concentrations of plant growth regulators (PGRs)
On 1st regeneration cycle, maximum 14.5 ± 0.31 and 9.7 ± 0.29 number of shoots were induced on Murashige and Skoog (MS) medium containing 2.5 mg/L BAP and 4.0 mg/L KIN respectively after 8 weeks of culture
Summary
Aloe vera (L.) Burm.f is an important industrial crop, which has enormous application in pharmaceutical, cosmetic and food industries. Present studies aim to develop a proficient methods of high-frequency true-to-type plantlet regeneration without intermediate callus phase for A. vera. Aim of our present studies is to develop a proficient and cost effective method for rapid and high frequency shoot multiplication and in vitro rooting of A. vera from rhizomatous stem explants. RAPD based detection of genetic polymorphism have been found successful application in describing somaclonal variability/ homogeneity of micropropagated individual of many plant species (Savita et al 2012; Paridaa et al 2013; Goswami et al 2013; Cheruvathur et al 2013; Kumar et al 2013; Haque and Ghosh 2013a). The present study was aimed at the following: (1) induction and regeneration of plants via direct shoot regeneration, (2) RAPD profiles analysis and (3) comparative cytogenetic assessment of two years old micropropagated plants and mother plant
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