As. Pac. J. Mol. Biol. & Biotech., Dec 1999 Vol. 7(1) : 53-60

As. Pac. J. Mol. Biol. & Biotech., June 2010 Vol. 18, 275-295

As. Pac. J. Mol. Biol. & Biotech., Apr 2014 Vol. 2, 191-198

Micropropagation of Tecomella undulata (Sm.) Seem. and genetic fidelity testing of in vitro raised plants

Suman Kumari and Narender Singh*

Department of Botany, Kurukshetra University, Kurukshetra-136119, India.

* Author for correspondence: Professor Narender Singh*
Department of Botany, Kurukshetra University, Kurukshetra-136119, India.
Email : This email address is being protected from spambots. You need JavaScript enabled to view it.

Abstract.

An efficient protocol has been developed for plant regeneration from shoot apex explants of Tecomella undulata. Explants were isolated from 2-3 year old plants, cultured on a shoot induction media, and fortified with different concentrations (0.25, 0.5, 1.0 or 2.0 mg l^-1) of auxins (NAA, IAA or 2, 4-D) and cytokinins (BAP or Kinetin). The greatest number of shoots (3.0) was obtained on the medium fortified with BAP (1.0 mg l^-1). The greatest root induction response (63.8%) was observed on MS half strength semisolid medium supplemented with 5.0 mg l^-1 NAA. The regenerated plantlets were acclimatized and transferred to soil for normal growth under field conditions and 60% survived. Random amplified polymorphic DNA markers were used to analyze the genetic fidelity of these in vitro raised plants. Out of the forty three RAPD decamers screened; only ten primers resulted in two to twelve scorable bands. These ten RAPD primers generated 51 amplicons in total, ranging from 200-2,500 bp in size with an average of 5.1 bands per primer. The amplification products were monomorphic in micropropagated plants and similar to the mother plants, confirming the genetic fidelity of in vitro raised plantlets and corroborating the fact that in vitro multiplication through direct organogenesis is the safest method for multiplying true to type plants

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As. Pac. J. Mol. Biol. & Biotech., Apr 2014 Vol. 2, 185-190

A simple in vitro method to study alkaline tolerance in alkaligrass (Puccinellia airoides) at high pH

Xue Zhang¹, Koji Nomura*¹, Katsuyoshi Shimizu² and Ke-Zhang Xu³

¹Graduate School of Life and Environmental Science, University of Tsukuba, Tennodai 1-1-1, Tsukuba, Ibaraki, 305-8572, Japan,
²Faculty of Life and Environmental Sciences, University of Tsukuba, Tennodai 1-1-1, Tsukuba, Ibaraki, 305-8572, Japan,
³Faculty of Agronomy, Jilin Agricultural University, Changchun 130118, Jilin Province, China.

* Author for correspondence: Koji Nomura
Graduate School of Life and Environmental Science, University of Tsukuba Tennodai 1-1-1, Tsukuba, Ibaraki, 305-8572, Japan.
Email : This email address is being protected from spambots. You need JavaScript enabled to view it. Tel.: +81-29-853-6692 Fax: +81-29-853-6692

Abstract.

Species belonging to genus Puccinellia are commonly called 'alkaligrass' because of their tolerance to saline-alkali environment. Although phyto-remediation of saline-alkali lands by planting alkaligrass is already achieved, its mechanisms of alkali tolerance need to be further comprehensively and thoroughly investigated. In a previous work, we revealed that alkaligrass had high tolerance to a high pH condition already during its germination and initial growth by growing seedlings in vitro. For further studies of alkali tolerance, an in vitro-culture system under a sterile condition is required. In a previous work, we cultured young alkaligrass seedlings on an agar medium to reveal their responses to a high pH stress. In the present work, we elaborated the in vitro method further and developed a convenient, efficient and reliable culture system. We compared the roots of the seedlings that were grown in the in vitro-culture system with those of field-grown mature grasses (pH 8.5). In spite of the differences in the growing environment and age, the morphological features of the roots under pH 8.5 were identical between the in vitro-grown and field-grown grasses. Here, we propose a simple in vitro-culture method of alkaligrass seedlings under a controlled condition. The alkaligrass seedlings in our in vitro-culture system are compatible to field-grown alkaligrass. Thus, our in vitro system is proved to be useful for further investigations, such as proteomics approach, of this plant.

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