N. Sarangi¹, A.B. Mandal^1*, A.K. Bandyopadhyay¹, T. Venugopal², S. Mathavan² and T.J. Pandian²

¹Biotechnology Laboratory, Central Agricultural Research Institute, Port Blair 744 101, India.
² School of Biological Sciences, Madurai Kamaraj University, Madurai, Tamil Nadu 625 014, India.

(Received 10 April 1999 / Accepted 19 October 1999)

Abstract.
The rainbow trout growth hormone (rtGH) gene under the transcription control of long terminal repeat of Rous sarcoma virus was successfully introduced into three Indian major carps (IMC) viz. rohu (Labeo rohita) catla (Catla catla) and mrigal (Cirrihinus mrigala), en route electroporated sperms. The present communication enumerates the results of an array of electroporation experiments aimed at standardising diverse variables like voltage, capacitance, resistance and pulse constant to achieve maximum transformation efficiency. Stable genomic integration of the intruded alien gene was demonstrated through slot blot hybridization in all the three species. Per cent transgenic individuals were largely varied in the different species, maximum of 25% being observed in rohu followed by 23 and 13% in mrigal and catla, respectively. This indicates that electroporated sperm mediated gene transfer may emerge as a convenient means in fish transgenic development specially in IMC. Optimisation of electroporation condition for rtGH transformation to produce transgenic IMC are discussed hereunder in detail.

Keywords: rtGH, electroporation, sperm mediation, Indian major carps

V.M. Kulkarni^1*, S.A. Ranade²,T.R. Ganapathi¹, P.Suprasanna¹, V.A. Bapat¹, K.K. Ussuf¹ and P.S. Rao¹

¹Nuclear Agriculture and Biotechnology Division,
Bhabha Atomic Research Center, Trombay, Mumbai 400 085, India.
'Plant Molecular Biology Division, National Botanical Research Institute, Lucknow 226 001, India

(Received 22 June 1999 / Accepted 190 October l999)

Abstract.
Banana, plantain and cooking banana (all belonging to the genus Musa) are crops of great importance for people in the developing countries and are rich sources of calories, several vitamins and several inorganic minerals. The cultivated bananas are derived from two diploid species, Musa acuminata (genome AA) and Musa balbisiana (genome BB) such that an entire range of triploid and tetraploid forms exist with varying degrees of either or both AA and BB genomes. The classification of banana germplasm has been confounded by the traditional practice of assigning local and vernacular names to the cultivars and varieties. The present study was carried out, in case of 29 varieties collected from different places in India, with the objective of distinguishing between the varieties at the molecular levels. Additionally, 1 genotype belonging to a closely related genus, namely, Ensete superbum was also included as an out group in the studies. The RAPD profiles of these 30 plants enabled distinction between the irradiated and unirradiated variety in an apparently dosage dependent manner. However, the cluster analysis did not group the varieites with any specific trend. Of all the varieties, 30 Gy irradiated Basrai and Williams were most similar to each other (SI=0.79) while the varieties Monthan and Chakkerkeli were the least similar (SI=0.22).

Keywords: Bananas, Musa, Ensete superbum, genetic variation, irradiation derived mutants, RAPD

G. Trejo Tapia¹, A. Jiminez Aparicio^1*, M. Rodriguez¹, G. Sepulveda¹, G. Salcedo¹, B. Martinez¹ , G. Gutierrez ² and A. De Jesus¹

¹Centro de Desarrollo de Productos Bioticos IPN, P.0. Box No. 24, Yautepec 62 731 Morelos Mexico.
²Escuela Nacional de Ciencias Biologicas IPN, Carpio y Plan de Ayala, s/n Col. Sto. Tomas,
Mexico 11340DF, Mexico

(Received 15 December 1998 /Accepted 6 October 1999)

Abstract.
To propose an alternative culture media for the production of betalains by cell suspensions of Beta vulgaris, the influence of the main macronutrients, nitrogen, phosphorous and calcium was investigated. Betalain production and cell growth were not affected significantly as a result of the nitrate/ ammonium ratios (3.02 to 12.23). The decrease from 24.72 to 12.36 mM on nitrate concentration and the suppression of ammonium from the standard B5 media did not affect betalain production and growth of the culture. Phosphate (0.5 to 1.5 mM) and calcium (from 0.5 to 1.5 mM) did not influenced significantly neither pigment synthesis nor growth. The elimination of calcium from the culture media resulted in a severe decrease on pigment production and cell growth and thus proving its importance for maintaining pigment production. On the other hand, a 100% increase in calcium concentration with respect to the concentration in B5, exerted a negative effect on the production. betacyanins/ betaxanthins (bc/bx) ratio was not modified with media changes. An alternative medium is proposed, containing no ammonium and 50% less nitrate, phosphate and calcium compared to standard B5 culture medium. The proposed modifications in the composition of the alternative medium, should reduce costs, not only when used for laboratory tests, but also when applied for bioreactor production system.

Keywords: Beta vulgaris, betalains, media culture, nitrogen, phosphate, calcium

S. Prasad¹, KS. Reddy² and N.Jawali^1*

¹Molecular Biology and Agriculture Division
²Nuclear Agriculture and Biotechnology Division
Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India

(Received 29 July 1999 / Accepted 10 October 1999)

Abstract.
In eukaryote genomes microsatellites are abundant and dispersed and they have gained importance as single locus markers because of their high level of polymorphism. The abundance of some tri and tetranuclcotide repeat microsatellites were surveyed in random amplified DNA fragments for the development of microsatellite markers for mungbean. Amplification products obtained from 39 arbitrary primers were screened. The data obtained on random amplified products hybridising microsatellites (RAHMs) show GGTT repeat RAHMs is the most abundant followed by CAA and CAC repeat RAHMs. The tetranucleoticle GCGT repeat RAHMs constitute nearly half as much as CAA repeat RAHMs, whilst GACA repeat RAHMs are absent. Some primer/ probe combinations were used for assessing RAHM polymorphism among 15 genotypes of mungbean. The polymorphic RAHMs (Random Amplified Microsatellite Polymorphism :RAMPO) were essentially dominant in nature (as in the case of RAPD).

Keywords: Arbitrary primers, microsatellites, mungbean, RAHM, RAMPO