Poonam Bhatia

Tissue Culture Studies of Tomato (Lycopersicon esculentum)

Tomato is a major vegetable crop that has achieved tremendous popularity over the last century. It is grown in almost every country of the world. Development of protocols for in vitro selection can provide new advances for the production of stress tolerant cultivars. Techniques have been optimised for the production of haploids and somatic hybrids. Attempts have also been made to transfer the higher regenerative ability of wild varieties to cultivated tomatoes. Although, some information is available on the morphogenesis of tomato, the techniques have not been developed to a level at which they can be utilised in large-scale multiplication of commercially important cultivars. The morphogenesis response seems to be highly dependent PGRs used in the media, which is again cultivar and genotypic specific. Somatic embryogenesis in tomato is still at its infancy, and efficient procedures for large-scale production via somatic embryogenesis are yet to be developed. Genetic stability of the tissue culture raised tomato plants also needs to be addressed. The use of a combination of molecular and conventional breeding techniques could be the option for the development of cultivars resistant to biotic and abiotic stresses. This paper reviews the advances made in various aspects of tissue culture in tomato. It also discusses the issues that still need to be addressed to utilise the full potential of plant tissue culture techniques in genetic improvement and mass propagation of tomato.

EFFECTS OF GENOTYPE, EXPLANT ORIENTATION, AND WOUNDING ON SHOOT REGENERATION IN TOMATO

SummaryEffects of genotype and explant orientation on shoot regeneration from cotyledonary explants of tomato were studied using 10 commercially important cultivars. The explant orientation affected shoot regeneration in all the tested genotypes. Cotyledons placed in abaxial (lower surface facing down) orientation consistently produced better shoot regenerative response and produced greater numbers and taller shoots compared to those inoculated in adaxial (upper surface facing down) orientation. Genotypic variation in terms of shoot regeneration response, number of shoots, and shoot height was apparent. Wounding of cotyledonary explants increased shoot regeneration response. However, shoot height was much lower in shoots regenerated from wounded explants compared to those that originated from intact cotyledons. Shoots produced from wounded cotyledons were abnormal in their form and structure.

In vitro propagation of Stackhousia tryonii Bailey (Stackhousiaceae): a rare and serpentine-endemic species of central Queensland, Australia

Stackhousia tryonii Bailey, a rare species whichhyperaccumulates nickel and with a potential to be exploited inphytoremediation/phytomining, is difficult to propagate via seeds. This studyinvestigated the development of a micropropagation protocol for the productionof large stocks of S. tryonii. Disinfested shoot tips andnodal buds were precultured on Gamborgs (B5) basal medium toobtain aseptic shoots for the optimisation of the protocol. 6-Benzyl aminopurine(BAP) at 1.0 mg l−1 produced the highest number ofshoots per explant in B5 medium. Comparison betweenB5 and MS media showed similar responses, but with marked influenceof BAP concentration on shoot numbers. Transfer of shoots from MS(multiplication) medium to MS medium supplemented with indole-3-acetic acid(IAA) and indole-3-butyric acid (IBA), individually or in combination, indicatedthat a combination of IAA and IBA (0.75 mg l−1each) is required to produce roots on young shoots (75%) compared to IBA(15–45%) or IAA (0–10%) alone. This study demonstrated that by usingthis protocol, a high multiplication rate (up to 18 shoots per explant) could be produced within 4 weeks, andthey can be readily hardened (98% survival) in a glasshouse by transplantingthem into a potting mixture of sand and perlite (4:1).

In vitro spore germination of the fern Schizaea dichotoma

Spores of Schizaea dichotoma were disinfested using sodium hypochlorite (NaOCl), streptomycin, or a combination of both streptomycin and NaOCl. Only the spores treated with streptomycin germinated. The highest germination (34%) was recorded in MS1/4 C compared to MS1/2, BB, or MM medium. The spore germination was not affected by the presence or absence of light or mycorrhizal fungi (Glomus clarum). The germinated spores developed up to five cells and failed to develop into a mature gametophyte.

Ex Vitro Rooting of Micropropagated Shoots of Stackhousia Tryonii

Micropropagated shoots of Stackhousia tryonii were exposed (individually or in combination) to indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), and 1-naphthalene acetic acid (NAA) at concentrations 1, 2 or 4 g dm−3 with the view to induce rooting under ex vitro conditions. The treated microshoots were grown in a mist room for four weeks and assessed for survival, rooting percentage, number of roots and root length. The results showed that IBA at 2 g dm−3 was most effective in inducing roots. Mixing of two or more auxins markedly reduced rooting percentage indicating antagonistic effects. The results demonstrated the potential of combining ex vitro rooting and hardening in one step, with view to reducing costs of multiplying plants via micropropagation.

Comparative Performance of Micropropagated and Seed-Grown Tomato Plants

Morphological, physiological, fruit yield and quality related traits were compared between the seed-grown and tissue-cultured plants of tomato (Lycopersicon esculentum Mill.) cv. Red Coat in a greenhouse. No significant differences were observed for any of the traits studied except for the number of leaves and branches, which were higher in the seed-grown plants than in tissue-cultured plants at the later stages of growth. No phenotypic abnormality of the tissue-cultured plants was observed suggesting that genetic fidelity of tissue cultured plants can be maintained if appropriate plant growth regulators are used with fewer member of subcultures in the multiplication medium.