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Indian Journal of Biotechnology

 

 

 

Special Issue on Plant Tissue Culture

 

ISSN:0972-5849

 

CODEN:IJBNAR 3(2) 145-322 (2004)

VOLUME 3

NUMBER 2

APRIL 2004

 

 

 CONTENTS

 

Micropropagation for quality propagule production in plantation forestry

159

Int. Cl.7 A 01 H 4/00, 5/00; C 12 N 15/00

 

R Yasodha, R Sumathi & K Gurumurthi

 

 

 

Tobacco (Nicotiana tabacum L.)—A model system for tissue culture interventions and genetic engineering

 

171

 Int. Cl.7 A 01 H 4/00, 5/00; A 61 K 35/74, 35/76, 39/002, 39/02, 39/12; C 12 N 15/00, 15/01, 15/05, 15/08, 15/09

 

 T R Ganapathi, P Suprasanna, P S Rao & V A Bapat

 

 

 

Isolated microspore culture of Brassica: An experimental tool for developmental studies and crop improvement

 

185

IPC Code: Int. Cl.7 A 01 H 1/08, 4/00,5/00; C 12 N 15/00, 15/01

 

Shashi B Babbar, Pradeep K Agarwal, Shalini Sahay & Sant S Bhojwani

 

 

 

Production of cloned trees of Populus deltoides through in vitro regeneration of shoots from leaf, stem and root explants and their field cultivation

 

203

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

H C Chaturvedi, A K Sharma, B Q Agha, M Jain & M Sharma

 

 

 

Studies on in vitro propagation of Himalayan cedar (Cedrus deodara) using zygotic embryos and stem segments

 

209

IPC Code: Int. Cl.7 A 01 H 4/00, 7/00

 

Sushma Tamta & L M S Palni

 

 

 

Micropropagation of an adult tree-Wrightia tinctoria

216

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

S D Purohit & G Kukda

 

 

 

Induced nucellar embryogenesis in vitro for clonal multiplication of Mangifera indica L. var. Ambalavi: A dwarfing rootstock

 

221

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

H C Chaturvedi, S Agnihotri, M Sharma, A K Sharma, M Jain, P Gupta, A Chourasia & N R Kidwai

 

 

 

An improved method of proliferation of proembryogenic calli of Mangifera indica L. var. Amrapali for scale-up of somatic embryo production

 

 

229

IPC Code: Int. Cl.7 a 01 H 4/00, 5/00

 

Hussain Ara, Uma Jaiswal & V S Jaiswal

 

 

 

In vitro cloning of female and male Carica papaya through tips of shoots and inflorescences

 

235

 IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

S Agnihotri, S K Singh, M Jain, M Sharma, A K Sharma & H C Chaturvedi

 

 

 

Cloning of adult trees of Jamun (Syzygium cuminii)

241

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

Vinod Rathore, N S Shekhawat, R P Singh, J S Rathore & H R Dagla

 

 

 

Embryogenesis and plant regeneration from mesocarp of Psidium guajava L. (guava)

246

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

 R Chandra, A Bajpai, Soni Gupta & R K Tiwari

 

 

 

Biotechnological interventions for genetic amelioration of Actinidia deliciosa var. deliciosa (kiwifruit) plant

 

249

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00; C 12 N 15/09

 

D R Sharma & Poonam Shirkot

 

 

 

In vitro cloning of Cajanus cajan var. Bahar through prolific shoot bud differentiation in leaf segments and production of fertile plants

 

258

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

M Jain & H C Chaturvedi

 

 

 

In vitro cloning of ornamental species of Dianthus

263

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

 Aparna Pareek, Archana Kantia & S L Kothari

 

 

 

Development of resin canals during somatic embryogenesis in callus cultures of Commiphora wightii

 

267

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

Sandeep Kumar, K C Sonie & K G Ramawat

 

 

 

In vivo and in vitro antimicrobial efficacy of Mimosa hamata

271

IPC Code: Int. Cl.7 A 01 H 5/00, A 01 N 65/00

 

S C Jain, R Jain & A J Vlietinck

 

 

 

Transgenic pants as bioreactors

274

IPC Code: Int. Cl.7 A 01 N 63/00; A 61 K 35/74, 35/76, 38/00, 39/02, 39/12;

C 12 N 15/00

 

 Arun K Sharma, Dewal Jani, C Raghunath & Akhilesh K Tyagi

 

 

 

Introgression of osmotin gene for creation of resistance against Alternaira blight by perturbation of cell cycle machinery

 

291

 IPC Code: Int. Cl.7 A 01 H 4/00, 5/00; C 12 N 15/09, 15/29

 

Gohar Taj, Anil Kumar, K C Bansal & G K Garg

 

 

Polyamine-ethylene nexus: A potential target for post-harvest biotechnology

299

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00; C 12 N 15/09

 

S Vinod Kumar & M V Rajam

 

 

 

In vitro germplasm preservation through regenerative excised root culture for conservation of phytodiversity

 

305

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

H C Chaturvedi, M Sharma, A K Sharma, M Jain, B Q Agha & P Gupta

 

 

 

Symposium Report

316

 

 

Instructions to Contributors

319

   
AUTHOR INDEX
 

 

 

Indian Journal of Biotechnology

 Vol. 3, April 2004, pp. 159-170

 

Micropropagation for quality propagule production in plantation forestry

R Yasodha, R Sumathi and K Gurumurthi*

 

Plantation forestry is the major source of raw material for industrial and domestic wood products and perpetually provides renewable energy, fiber and timber. Successful plantation forestry is dependent on effective research and development leading to technological advances. In hard wood plantation establishment, micropropagation is preferred for the production of quality planting stock because of its very high multiplication rate with instant silvicultural gains. Integration of micropropagation in tree improvement is essential for the replication of improved genetic material and rapid release of quality propagules. Further, in vitro propagation is also required for rejuvenating productive clones, which are difficult to root; rescuing of important mature individuals and natural hybrids; and producing stock plants for clone bank establishment and conserving the important germplasm. Thus, the approaches for micropropagation should be species oriented to fit into the existing strategies of tree improvement. In this paper, the recent advances made towards the production of quality plantlets through micropropagation of hardwood tree species, such as eucalypts, acacia, teak and bamboos, are presented. Further, the strategies for the multiplication of elite genotypes and the use of microproagules as planting stock in plantation forestry are discussed.

 

Keywords: micropropagation, tree improvement, plantation forestry, eucalypts, acacia, teak, bamboos

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00; C 12 N 15/00

 

 

 

Indian Journal of Biotechnology

 Vol 3, April 2004, pp 171-184

 

Tobacco (Nicotiana tabacum L.)—A model system for tissue culture interventions and genetic engineering

T R Ganapathi1, P Suprasanna1, P S Rao2 and V A Bapat1*

 

Tobacco (Nicotiana tabacum L.) has become a model system for tissue culture and genetic engineering over the past several decades and continues to remain the ‘Cinderella of Plant Biotechnology’. An in vitro culture medium (Murashige and Skoog, 1962), based on the studies with tobacco tissue cultures, has now been widely used as culture medium formulation for hundreds of plant species. Studies with tobacco tissue culture have shed light on the control of in vitro growth and differentiation. Further, induction of haploids, microspore derived embryos and selection of mutant cell lines, have been achieved successfully. Tobacco has also been employed for the culture and fusion of plant protoplasts, providing invaluable information on way to explore the potential of somatic hybridization in other crops. Optimization of genetic transformation, using Agrobacterium tumefaciens and A. rhizogenes, has been central to the cascade of advances in the area of transgenic plants. Developments in the field of molecular farming for the expression and/or production of recombinant proteins, vaccines and antibodies are gaining significance for industrial use and human healthcare.

 

Keywords:   genetic transformation, molecular farming, plant biotechnology, plant cell and tissue culture, recombinant proteins, tobacco

IPC Code:   Int. Cl.7 A 01 H 4/00, 5/00, A 61 K 35/74, 35/76, 39/002, 39/02, 39/12; C 12 N 15/00, 15/01, 15/05, 15/08, 15/09

 

 

 

 

Indian Journal of Biotechnology

 Vol. 3, April 2004, pp 185-202

 

Isolated microspore culture of Brassica:
An experimental tool for developmental studies
and crop improvement

 Shashi B Babbar1, Pradeep K Agarwal1, Shalini Sahay1 and Sant S Bhojwani2*

 

The technique of isolated microspore culture for the production of haploid plants is well standardized for many species of Brassica, some of which are important oilseed or vegetable crops. It offers an excellent experimental system for improve-ment of Brassica crops and study of cell differentiation during shift from gametophytic to sporophytic development. The haploids raised from microspores are of tremendous use in genetic improvement of brassicas through breeding, mutant induction and selection, and genetic engineering. The present review describes current status of the technology, the factors influencing the success of microspore culture, structural, biochemical and molecular changes occurring in the microspores triggered towards sporophytic development, and the progress made in practical applications of the microspore-derived plants.

 

Keywords: Brassica, isolated microspore culture, anther culture, haploids, crop improvement, diplodization, mutants, genetic transformation, germplasm storage

IPC Codes: Int. Cl.7 A 01 H 1/08, 4/00, 5/00; C 12 N 15/00, 15/01

 

 

 

Indian Journal of Biotechnology

 Vol 2, April 2004, pp 203-208

 

Production of cloned trees of Populus deltoides through in vitro regeneration of shoots from leaf, stem and
root explants and their field cultivation

 H C Chaturvedi*, A K Sharma, B Q Agha, M Jain and M Sharma

 

Amongst a large number of Populus species, micropropagation of P. deltoides has not been possible. The authors now report a reproducible protocol for mass production of cloned plants of 2 important clones of P. deltoides, viz., G3 and G48 through induced shoot bud differentiation in the in vitro cultured segments of leaf, stem and root of adult tree origin. Establishment of experimental plants in aseptic culture was possible only when nodal stem segments from fresh flush of growth were initially cultured in a liquid nutrient medium, a modification of Murashige and Skoog (1962) medium (MS) supplemented with 0.25 mg l-1 each of BAP and IAA and 15 mg l-1 AdS. Axillary shoot growth from nodal stem segments was sustained in an agarified medium of the same composition following at least 2 subcultures in the liquid medium. Rate of multiplication got enhanced many-fold when the segments of leaf, stem and root of regenerated shoots were induced to differentiate multiple shoot buds in the same medium instead of obtaining a single plantlet per regenerated shoot. The maximum number of shoot buds, i.e., 28.8 was differentiated in leaf segments followed by 22.7 and 7.8 in stem and root segments, respectively. However, the regenerated shoots appeared similar both morphologically and in vigour irrespective of their origin from any of the explants. Explants with differentiated shoot buds on being subcultured in the same morphogenic medium repeatedly produced crops of proliferated shoots. Developed isolated shoots, measuring about 3 cm in length, rooted 100% in 20-25 days, in a liquid rooting medium having 0.25 mg l-1 IAA. The corresponding cultures on agarified medium of the same composition not only rooted poorly, but also necrosed and died. The rooted shoots required precision during their hardening in a purely inorganic salt solution following the procedure developed earlier. It took about.12 months to obtain hardened plants of about 30 cm in height in potted soil from the time of establishment of aseptic cultures using nodal stem segments of an adult tree. The properly hardened plants survived more than 80% on their transplantation to soil and grew luxuriantly under field conditions. The in vitro-raised trees attained an average height of 5 m with straight boles of an average diameter of 12 cm in 2 yrs.

 

KeywordsPopulus deltoides, in vitro cloning, leaf segments, nodal stem segments, root segments, shoot bud differentiation, stem segments

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

 

 

Indian Journal of Biotechnology

 Vol. 3, April 2004, pp. 209-215

 

Studies on in vitro propagation of Himalayan cedar (Cedrus deodara) using
zygotic embryos and stem segments

Sushma Tamta* and L M S Palni

 

Adventitious bud formation was examined in zygotic embryos (decoated seeds) of Cedrus deodara, planted in the normal position, using four different media variously supplemented with or without plant growth regulators (PGRs). Following incubation for 7, 14, 21 and 28 days on PGR containing media, the embryos were transferred to respective PGR-free basal media. While embryo swelling was observed in all cases, normal germination was observed only in embryos cultured initially in PGR-free media. Best response, in terms of bud formation (93%), was observed in embryos first cultured on LP medium containing BA and Kinetin (1.0 mM each) for 28 days and then transferred to LP medium without any PGR. While the mean number of shoots formed per cultured embryo was also highest (34%) in this combination, these shoots eventually dried when excised and sub-cultured for further multiplication. Success was, however, obtained when the embryos were initially planted in an inverted position on LP medium containing 20.0 mM BA for 10 days, and then shifted, again in the inverted position, on the same medium without BA. Although the mean number of shoots formed was low (only 2.5 shoots per embryo), these could be multiplied and elongated. The microshoots (2.0-3.0 cm) thus obtained could be rooted (100%) using ½ strength, PGR-free, LP medium, hardened and established in pots. Besides excised embryos, stem segments were also used as explants. While only a quarter of explants could be established in culture, in contamination-free condition, good sprouting response (3-4 buds per segment) was obtained in certain PGR combinations. The sprouted buds developed into shoots on LP medium containing 5.0 mM BA; further subculturing of excised shoots for multiplication, however, resulted in drying up of shoots. Further work is required to overcome this difficulty.

 

Keywords: Cedrus deodara, conifer, deodar, micropropagation, inverted embryo technique, stem segments, rooting

IPC Code: Int Cl.7 A 01 H 4/00, 7/00

 

 

 

Indian Journal of Biotechnology

 Vol. 3, April 2004, pp. 216-220

 

Micropropagation of an adult tree-Wrightia tinctoria

S D Purohit1* and G Kukda2

 

Multiple shoots were induced in vitro from nodal shoot segments of a 30-yr-old plus tree having enhanced axillary branching. Initiation of culture was strongly influenced by the nature of explant and the season of harvesting. Nodal segments (91%) from young lateral branches produced an average of 5 shoots per node in 3 weeks on agar solidified MS medium supplemented with 2.0 mg l-1 benzylaminopurine (BAP). After establishment of cultures and initiation of shoot buds, a cluster of shoots including mother explant, was transferred to medium containing a lower concentration of BAP (1.0 mg l-1). A three-fold rate of shoot multiplication during every subculture of 3 weeks was achieved. Nodal segments from in vitro raised shoots were also used to initiate a new culture cycle. The shoots could be multiplied for at least 24 months without loss of vigour. The shoots (71%) were successfully rooted when their lower ends were dipped in pre-autoclaved indole-3-butyric acid solution (100 mg l-1) for 10 min followed by their implantation on modified MS medium (major salts reduced to 1/4 strength) containing 200 mg l-1 activated charcoal. Out of 2000 plants, 1600 (80%) were successfully hardened under greenhouse conditions and more than 200 have been planted out in soil where they are growing well.

 

Key words: clonal propagation, Wrightia tinctoria, tissue culture, micropropagation

IPC Code: Int Cl.7 A 0 1 H 4/00, 5/00

 

 

 

Indian Journal of Biotechnology

 Vol 3, April 2004, pp. 221-228

 

Induced nucellar embryogenesis in vitro for clonal multiplication of Mangifera
indica
L. var. Ambalavi: A dwarfing rootstock

H C Chaturvedi*, S Agnihotri, M Sharma, A K Sharma, M Jain, P Gupta,

A Chourasia and N R Kidwai

 

Nucellar embryogenesis was induced in Mangifera indica L. var. Ambalavi, a monoembryonic dwarfing rootstock. Nucellar tissue of young fruits of 3 developmental stages and measuring about. 2.5, 4 and 5 cm in length, responded differently to the same treatments of agarified nutrient medium used for induction of embryogenesis. Whilst 0.25 mg l-l BAP along with 1 mg l-l NAA was effective to induce embryogenesis in nucellus of youngest fruits, 0.5 mg l-1 2iP alone was sufficient for nucellus of older fruits. However, all the differentiated embryos proliferated in a single treatment comprising 0.15 mg l-1 each of BAP and 2iP and 0.5 mg l-l IAA, albeit with a different basal medium. Size of cotyledonary nucellar embryos ranging between 1.5 to 2 cm in length was necessary for their further development, maturation, germination (visible plumule and developed root) and convertibility (plantlet formation). Embryos of desirable size required the liquid state of the medium supplemented with 0.01 mg l-1 ABA, 0.1 mg l-1 IAA and 100 mg l-1 PEG (M. Wt. 400), while the nitrogen content was also low for their near-synchronized development, maturation, germination and convertibility.  In the optimum treatment, most of the embryos showed apparently normal development, of which 78.4% matured and 40.2% germinated, while 35.6% embryos produced plantlets. Such a high percentage of convertibility of nucellar embryos has not been obtained earlier. The hardened in vitro-raised plantlets survived ex vitro when first transplanted to Soilrite followed by their transfer to the garden soil. Furthermore, adventitiously rooted nucellar plantlets gave better transplant success, i.e., about. 70% than those with their initial root system, where it was about. 50%. The former also survived longer than the latter, i.e., beyond 4 months.

 

Keywords: Ambalavi, cloning, dwarfing rootstock, Mangifera indica, monoembryonic, nucellar embryogenesis, rooted         shoots

IPC Code: Int Cl.7 A 01 H  4/00, 5/00

 

 

 

Indian Journal of Biotechnology

 Vol. 3, April 2004, pp 229-234

 

An improved method of proliferation of proembryogenic calli of Mangifera indica L. var. Amrapali for scale-up of somatic embryo production

Hussain Ara, Uma Jaiswal and V S Jaiswal*

 

Rapid enhancement of proliferation of PECs (proembryogenic calli), obtained from the excised nucellus tissue of var. Amrapali, with high efficiency somatic embryo production has been achieved by manipulating the medium with different growth regulators. Among the growth regulators tested, 1.0 mg/l 2,4-D (2,4-dichlorophenoxy acetic acid) and 1.0 mg/l NAA (a-naphthalene acetic acid), either alone or with 1.0 mg/l Kn (kinetin), stimulated proliferation of the PEC in both the liquid and solid medium; although, it was more profuse (<5 times) in the liquid medium. Depending upon the physical state and growth regulator(s) supplemented to the medium, the PECs appeared in three morphotypes. Type–III PEC, which was in the form of fine suspensions of dispersible and highly proliferating single cells and small cell aggregates, showed greater potentially for differentiation of somatic embryos in both states of the medium; however, in liquid medium the globular somatic embryos dedifferentiated and callused soon after their differentiation. In contrast, on the semi-solid medium these globular somatic embryos successfully developed into cotyledonary-stage. Type-III PEC (10 mg) had the capacity to produce up to 350 somatic embryos on semi-solid medium. Such somatic embryos matured and grew into plantlets. The method offers a possibility of rapid multiplication in the mango var. Amrapali.

 

Keywords: excised nucellus, growth regulators, Mangifera indica L., proembryogenic callus (PEC), somatic embryogenesis

IPC Code: Int. Cl7. A 01 H 4/00, 5/00

 

 

 

Indian Journal of Biotechnology

 Vol 3, April 2004, pp 235-240

 

In vitro cloning of female and male Carica papaya through tips of shoots and inflorescences

S Agnihotri*, S K Singh, M Jain, M Sharma, A K Sharma and H C Chaturvedi

 

Raising cultures of proliferating shoots of female and male plants of Carica papaya through shoot apices taken from mature plants was difficult because of high incidence of endogenous bacterial contamination. Whilst they were easily raised through culture of young inflorescences tips of female and male plants. There was no difference in the requirement of nutrients and growth regulators for proliferation of shoots raised from shoot tips or inflorescence tips, but at the initial stage, the different explants differed in their requirement of growth regulators for induction of shoot bud differentiation. BM1, a modification of MS medium was used as the basal medium in all the cases. There was a pronounced callusing tendency in tender shoots initially used for rooting, for which well-developed shoots (< 4 cm long) were found suitable. Rooting was achieved in developed shoots through a 4-step procedure: Step 1-An initial pulse treatment with a high concentration of IBA (10 mg l-1) using BM3 medium, a modified Knop medium with trace elements and disodium-ferric-ethylene-diaminetetra-acetate (Na-Fe-EDTA) of MS medium and 2% sucrose for 24 h; Step 2- Their subculture in medium BM2 differing from BM1 in having 50 mg l-1 m-inositol and supplemented with IAA (0.25 mg l-1) along with AdS (15 mg l-1) and 2% sucrose for 7 days; Step 3- Roots at the cut end of about 95% shoots were visible, after about 10 days, in the same medium as used in Step 1, but having supplements of 3 vitamins, 2 amino acids and 0.25 mg l-1 IAA, while sucrose was removed; and Step 4- The just rooted shoots when finally transferred to the semi-sterilized moist Soilrite contained in culture tubes, formed healthy roots without intervening callusing, while the shoots also remained healthy. Such plantlets when ultimately transplanted along with Soilrite plugs to the potted soil showed about 80% transplant success. In vitro-raised plants appeared normal and fruited under field conditions after about 6 months of ex vitro growth.

 

Keywords: Dioecious, endogenous infection, ex vitro growth, intervening callusing, in vitro rooting, long-term culture, shoot proliferation

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

 

Indian Journal of Biotechnology

 Vol. 3, April 2004, pp. 241-245

Cloning of adult trrees of Jamun (Syzygium cuminii)

Vinod Rathore, N S Shekhawat*, R P Singh, J S Rathore and H R Dagla

 

For cloning of adult tree(s) of Jamun (Syzygium cuminii), explants were harvested from rejuvenated shoot sprouts produced by lopped tree(s). Multiple shoots regenerated by activation of axillary meristems of the explants on MS + 9.0 µM BAP + growth additives. Shoots were proliferated in culture by (i) repeated transfer of original explants, and (ii) subculture of shoots on amended medium with combination of BAP (4.50 µM) + kinetin (2.25 µM). The cloned shoots were rooted (i) in vitro on half-strength MS + 0.1% activated charcoal supplemented with IBA (10 µM) or NAA (15.0 µM), and (b) ex vitro by pulse treatment with 2.50 mM of IBA. The shoots root 100% under ex vitro conditions. The cloned treelets were hardened and pot transferred. The ex vitro rooting of cloned shoots was highly effective; it saved time and resources, and can be used for cloning of Jamun.

 

Keywords: Cloning, Jamun, mature tree, nodal shoots, Syzygium cuminii

IPC Code: Int.Cl.7 A 01 H 4/00, 5/00

 

 

 

Indian Journal of Biotechnology

 Vol. 3, April 2004, pp. 246-248

 

Embryogenesis and plant regeneration from mesocarp of Psidium guajava L. (guava)

R Chandra*, A Bajpai, Soni Gupta and R K Tiwari

 

A protocol has been developed for the induction, maturation and germination of somatic embryos from mesocarp tissue of Psidium guava L. (guava). Explants were cultured in modified MS medium fortified with 2, 4-D (2.0 mg/l), ascorbic acid (100 mg/l), L-glutamine (400 mg/l) and sucrose (6%). Embryogenic proliferating tissue was induced, which was translucent, mucilaginous and differentiated into many small somatic embryos. The somatic embryos were retained in the same medium, where simultaneously differentiation of new somatic embryos and their conversion into plantlets was observed. Thus, the embryogenesis in guava could be perpetuated, which would be used in future for carrying out cellular selection against wilt causing organism.

 

Keywords: Psidium guajava, guava, mesocarp, somatic embryogenesis, repetitive embryogenesis

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

 

 

Indian Journal of Biotechnology

 Vol 3, April 2004, pp 249-257.

 

Biotechnological interventions for
genetic amelioration of Actinidia deliciosa var. deliciosa (kiwifruit) plant

D R Sharma* and Poonam Shirkot

 

Kiwifruit (Actinidia deliciosa var deliciosa) is a recent introduction among the promising fruit crops. Popularity of its nutritious fruit berries has created a substantial demand for quality planting material, which needs to be planted in certain proportions of female and male plants for Actinidia is dioecious. The large-scale propagation of planting material can be achieved through micropropagation; whereas, it’s quality and clonal nature need to be ensured, preferably, through molecular markers. With further popularity of this fruit crop, tailor-made genotypes will be required, which can be made possible through biotechnological interventions. As A. deliciosa is limited in its adaptation to a very narrow range of agro-climatic conditions, there is also a need to develop culivars endowed with traits of wider adaptation. So that this fruit crop can be cultivated in larger range of geographical areas. Development of micropropagation protocols, identification of gender at seedling stage, identification of molecular markers to test genetic/clonal fidelity and incorporation of genes for genetic amelioration are the issues that need immediate consideration. Some of these issues have been addressed in the present paper.

 

Keywords: Actinidia, conservation, genetic transformation, in vitro techniques, kiwifruit, molecular markers

 

 

 

Indian Journal of Biotechnology

 Vol. 3, April 2004, pp. 258-262

 

In vitro cloning of Cajanus cajan var. Bahar through prolific shoot bud differentiation in leaf segments and production of fertile plants

M Jain and H C Chaturvedi*

 

Prolific differentiation of shoot buds was induced in the interveinal leaf lamina explants (5 mm 2 mm) excised from first pair of leaves of aseptically grown plants of Cajanus cajan var. Bahar. An explant cultured on filter paper bridge in a nutrient liquid medium supplemented with 0.25 mg l-1 each of BAP and Kn along with 0.05 mg l-1 IAA and 40 mg l-1 AdS, produced an av. 50 shoot buds within 45 days, after subjected to a pulse treatment comprising 2 mg l-1 each of BAP and Kn with 1 mg l-1 IAA for three days given under the same culture conditions. Development of normal shoots and simultaneous differentiation of new shoot buds were obtained by subculture of explants with differentiated shoot buds on agarified nutrient medium mainly in the presence of 0.01 mg l-1 each of BAP and IAA along with 10 mg l-1 CCC, where even more than 100 shoot buds were formed in the best cultures. Developed shoots rooted 100% in 0.25 mg l-1 of any of the three auxins used, viz, IAA, IBA and NAA, while the clonal plantlets gave 90% transplant success. In vitro-raised plants, when transplanted to field in mid-October, grew normally and produced viable seeds by late-March. The seeds, so produced, germinated 100% in vitro and gave rise to normally growing plants in soil. The protocol for cloning of this commercially important variety also offers an excellent system for its genetic transformation in having certain outstanding features, which include simultaneous differentiation of a great number of shoot meristems in a uniform leaf tissue explant and development of virtually all of them into normal fertile plants.

 

Keywords: Bahar, Cajanus cajan, fertile plants, interveinal leaf lamina, in vitro cloning, shoot bud differentiation

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

 

 

Indian Journal of Biotechnology

 Vol 3, April 2004, pp 263-266

 

In vitro cloning of ornamental species of Dianthus 

Aparna Pareek, Archana Kantia and S L Kothari*

 

Dianthus caryophyllus, D. chinensis and D. barbatus were micropropagated from shoot tips and nodal segments, which were cultured on MS medium supplemented with BAP and NAA. Best response in terms of multiple shoot formation was observed on MS medium with NAA (0.5 mg/l) + BAP (1 mg/l). In four weeks 10-15 shoots were formed. Every two weeks, inoculum of 6-8 shoots was transferred to a fresh medium for further shoot differentiation. Rooting of in vitro shoots was achieved on half strength, MS + IAA (0.5 mg/l). Plantlets that were transferred to pots matured and flowered and resembled seed grown plants Variation of any sort could be avoided as regeneration of shoots was obtained without intervening callusing.

 

Keywords: Dianthus caryophyllus, Dianthus chinensis, Dianthus barbatus, shoot tip, nodal segments, shoot bud  proliferation

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

 

 

Indian Journal of Biotechnology

 Vol. 3, April 2004, pp 267-270

 

Development of resin canals during somatic embryogenesis in callus cultures of Commiphora wightii

Sandeep Kumar, K C Sonie and K G Ramawat*

 

Formation of resin canal in somatic embryos of Commiphora wightii (Arnott.) Bhandari has been reported. Resin canal formation is a prerequisite for obtaining resin canal based secondary metabolites and resin canal formation in callus cultures is not known. Resin canals were observed in torpedo shaped and cotyledonary stage embryos. Early stages were devoid of it. Resin canals formed in somatic embryos were comparable with those formed in the stem. This provides large quantities of aseptic resin canals in organized structures, which can be used for the production of guggulsterones through biotechnological methods relieving pressure on natural resources.

 

Keywords: Commiphora wightii; somatic embryogenesis; resin canals; callus cultures

IPC Code: Int Cl.7 A 01 H 4/00, 5/00

 

 

 

Indian Journal of Biotechnology

 Vol 3, April 2004, pp 271-273

 

In vivo and in vitro antimicrobial efficacy of
Mimosa hamata

S C Jain1*, R Jain2 and A J Vlietinck3

 

Mimosa hamata Willd. (Mimosaceae) has demonstrated pronounced bioefficacy against the selected bacteria (Escherichia coli, Klebsiella aerogenes, Proteus vulgaris, Staphylococcus aureus and Pseudomonas aeruginosa), fungi (Aspergillus flavus, Fusarium moniliforme and Rhizoctonia bataticola) and viruses (Herpes simplex, Poliomyelitis and Vesicular stomatitis).

 

Keywords: Mimosa hamata, cell cultures, antibacterial, antifungal, antiviral efficacies

IPC Code: Int. Cl.7 A 01 N 65/00, A 01 H 5/00

 

 

 

 

Indian Journal of Biotechnology

 Vol 3, April 2004, pp 274-290

 

Transgenic plants as bioreactors

Arun K Sharma, Dewal Jani, C Raghunath and Akhilesh K Tyagi*

 

Recent advances in molecular and cellular biology have led to the development of technology to engineer plants that are capable of producing a wide variety of products. These include products for pharmaceutical applications, like vaccine antigens, antibodies, antibody-derived fragments and other therapeutic proteins. It is possible to alter endogenous metabolic pathways to obtain new products, like plastics, or to confer novel traits, such as improved nutritional value.

 

Keywords: antibody, antigen, bio-degradable plastics, bioreactors, nutrition, plantibody, transgenic plants

IPC Code: Int.Cl.7 A 01 N 63/00; A 61 K 35/74, 35/76, 38/00, 39/02, 39/12; C 12 N 15/00

 

 

 

Indian Journal of Biotechnology

 Vol. 3, April 2004, pp. 291-298

 

Introgression of osmotin gene for creation of resistance against Alternaira blight by perturbation of cell cycle machinery

Gohar Taj1*, Anil Kumar1, K C Bansal2 and G K Garg1

 

Pathogenesis related protein, osmotin, when introgressed into Brassica juncea plants, provided tolerance to fungal attack. Osmotin is thought to influence the signal transduction pathway. Possible influence of osmotin gene transfer on the cell cycle and cell death pathways was investigated using transformed B. juncea calli as model system. It was observed that partially purified Alternaria toxin inhibited the growth of non-transformed calli; whereas, transformed calli resisted the effect of toxin, which appears to counteract the inhibitory effects of phytotoxins. The toxin also initiates the programmed cell death (PCD) in the calli. The toxin increased the expression of P53 and caspase-like proteins in the non-transformed calli and suppressed the expression of cyclin B and CDC proteins. It was further supported by the fragmentation of DNA in response to toxin treatment. However, the transformed calli, when challenged with toxin, suppressed the expression of P53, while caspase 1 was not affected. On the basis these results, it can be concluded that osmotin tampers the P53-mediated PCD pathway; which, although, could not provide the resistance but delayed the appearance of disease symptoms.

 

Keywords: Alternaira blight, cell cycle, osmotin

IPC Code:Int. Cl.7 A 01 H 4/00, 5/00; C 12 N 5/09, 15/29

 

 

 

Indian Journal of Biotechnology

 Vol 3, April 2004, pp. 299-304

 

Polyamine-ethylene nexus: A potential target for post-harvest biotechnology

S Vinod Kumar and M V Rajam*

 

Post-harvest biotechnology is a fast growing field of plant biotechnology. Achievements were made in the field to delay or prevent fruit ripening and softening by genetic manipulation of ethylene and cell wall metabolism. Despite the development of transgenic plants with the desired traits, the technology has not achieved the pace due to drawbacks of the existing strategies to effectively control the developmental processes following harvest. Here the authors analyse various strategies of post-harvest biotechnology and prospects, giving special emphasis to the metabolic interactions of pathways related to post-harvest characteristics¾ those of polyamine and ethylene¾ and the engineering of polyamine metabolism as an alternate strategy for post-harvest biotechnology by carefully manipulating the delicate metabolic balance and the benefits of the newly proposed strategy over the currently used ones.

 

Keywords:   polyamines, ethylene, fruit ripening, senescence, transgenic plants

IPC Code:   Int. Cl.7 A 01 H 4/00, 5/00; C 12 N 15/09

 

 

 

Indian Journal of Biotechnology

 Vol. 3, April 2004, pp. 305-315

 

In vitro germplasm preservation through regenerative excised root culture for conservation of phytodiversity

H C Chaturvedi*, M Sharma, A K Sharma, M Jain, B Q Agha and P Gupta

 

In vitro preservation of germplasm becomes imperative for those plant species in which application of conventional methods is infructuous. Of the 4 methods of in vitro preservation of germplasm, viz., cryopreservation, slow growth shoot culture, normal growth culture and regenerative excised root culture, the last method has some advantages over the others in case of certain plant species, particularly the palms. Its main advantages are: Practicability and low cost because of no requirement of agar-agar, of light as well as of strict maintenance of temperature besides easy exchange of germplasm across the International boundaries without damage in transit. However, all these 4 methods have certain shortcomings, which necessitate adoption of a composite approach involving all of them to achieve the main goal of conservation of phytodiversity. In general, durations of germplasm preservation in vitro through cryopreservation and slow growth shoot culture have ranged from a few weeks to about 2 yrs depending on the plant species concerned. In certain plant species, the normal growth culture had afforded germplasm preservation for considerably long periods of time, viz., more than 27 yrs in Dioscorea floribunda and D. deltoidea and 32 yrs in Citrus grandis, albeit with a danger of cultures getting infected during frequent subcultures. In contrast, the duration of germplasm preservation through regenerative excised root culture ranged from 6 to 24 yrs. The method of regenerative excised root culture had been demonstrated to preserve germplasm of a number of plant species, including herbaceous annuals and perennials and trees, viz., Solanum khasianum (spiny and spineless), S. torvum, S. surattense, Atropa belladonna, Kalanchöe fedtschenkoi, Rauvolfia serpentina, Populus deltoides and Dalbergia latifolia. A similar possibility existed in case of Shorea robusta, Cocos nucifera and Elaeis guineensis, roots of which though established in long-term culture, the regenerant differentiation in their explants was most sporadic. For establishing excised root cultures of different plant species, modifications of various nutrient media, viz., Murashige and Skoog (1962), Street (1954), Street and McGregor (1952) and White (1943) were used in liquid state, while for inducing regenerant differentiation, some other modifications of the same media except of White as well as of Schenk and Hildebrandt (1972) medium were employed, using different cytokinins, viz., BAP, 2iP, Z and TDZ with auxins, viz., IAA, NAA and 2, 4-D and also certain growth inhibitors/retardants, viz., ABA, CCC and ancymidol as also a polyamine, putrescine. In all cases, agarified media were used except in R. serpentina, which required the liquid state of medium for regenerant differentiation, while in S. khasianum and A. belladonna, caulogenesis took place in agarified morphogenic medium and embryogenesis in the liquid state of medium. Further, whilst in R. serpentina and A. belladonna plantlets were produced via embryogenesis, in rest of the plant species through caulogenesis. Plants regenerated through long-term excised root culture were true-to-type, which was substantiated by tracing their origin from the pericycle tissue of root explants. The root-regenerated plants fared well under field conditions save A. belladonna, plants of which could be grown in potted soil only in controlled physical
conditions.

 

Keywords: Caulogenesis, conservation, embryogenesis, excised root culture, germplasm preservation, phytodiversity, regenerants from pericycle, root-regenerated plants

IPC Code: Int. Cl.7 A01 H 4/00, 5/00

 

 

 

 

EXPLANATORY NOTE ON IPC Int. Cl.7

 

The field of biotechnology has seen exponential growth in recent times. It attracted the attention of R&D institutes, who have made significant achievements/breakthroughs in the fields of agriculture, health and medicine, environment and so on, and consequently resulted in granting of hundreds of patents by patent authorities the world over. It has also thus aroused concerns in the above areas on ethical issues as well as profit making/sharing. In this context, patents, their classification, access to patent databases, prior art, etc. have assumed great significance. In biotechnology, these issues as such are more crucial since it is completely a field of applied nature.

 

From the present issue, Indian Journal of Biotechnology has decided to assign IPC codes to each article for creating more awareness for its readers, and facilitating the search of patent examiners as well as establishing prior art. The subject matter is briefly explained below.

 

The International Patent Classification, which is commonly referred to as the IPC, is based on an international multi-lateral treaty administered by World Intellectual Property Organization, Geneva, Switzerland.

 

The Classification is indispensable for the retrieval of patent documents in the search for "prior art." Such retrieval is needed by patent-issuing authorities, potential inventors, research and development units, and others concerned with the application or development of technology.

 

In order to keep the IPC up to date, it is continuously revised and a new edition is published every five years. The current (seventh) edition has entered into force on January 1, 2000.

 

The IPC is a hierarchical classification system comprising sections, classes, subclasses and groups (main groups and subgroups). The seventh edition of the IPC consists of 8 sections, 120 classes, 628 subclasses and approximately 69,000 groups.

 

Every subdivision of the IPC is indicated by a symbol and has a title. The IPC divides all technological fields into eight sections designated by one of the capital letters A through H. Each section is subdivided into classes. In turn, each class contains one or several subclasses, for example, A 01 B.

 

Each subclass is broken down into subdivisions referred to as "groups", which are either main groups or subgroups. Main group symbols consist of the subclass symbol followed by a one-to three-digit number, the oblique stroke and the number 00, for example, A 01 B 1/00. Subgroups form subdivisions under the main groups. Each subgroup symbol includes the subclass symbol followed by the one-to three-digit number of its main group, the oblique stroke and a number of at least two digits other than 00, for example, A 01 B 1/02.

 

For further information, please refer to: www.wipo.int

 

 

AUTHOR INDEX

 

Agarwal, P K

185

Jaiswal, U

229

Sahay, S

185

Agha, B Q

203, 305

Jaiswal, V S

229

Sharma, A K

203, 221, 235,305

Agnihotri, S

221, 235

Jain, M

203, 221, 235,258, 305

Sharma, Arun K

274

Ara, H

229

Jani, D

274

Sharma, D R

249

 

 

Jain R

271

Sharma, M

203, 221, 235, 305

Babbar, S B

185

Jain S C

271

Shekhawat, N S

241

Bajpai, A

246

 

 

Shirkot, P

249

Bansal, K C

291

Kantia, A

263

Singh, R P

241

Bapat, V A

171

Kidwai, N R

221

Singh, S K

235

Bhojwani, S S

185

Kothari, S L

263

Sonie, K C

267

 

 

Kukda, G

216

Sumathi, R

159

Chandra, R

246

Kumar, A

291

Suprasanna, P

171

Chaturvedi, H C

203, 221, 235, 258, 305

Kumar, S

267

 

 

Chourasia, A

221

 

 

Taj, G

291

 

 

Palni, L M S

209

Tamta, S

209

Dagla, H R

241

Pareek, A

263

Tiwari, R K

246

 

 

Purohit, S D

216

Tyagi, A K

274

Ganapathi, T R

171

 

 

 

 

Garg, G K

291

Raghunath, C

274

Vinod Kumar, S

299

Gupta, P

221, 305

Rajam, M V

299

Vlietinck A J

271

Gupta, S

246

Ramawat, K G

267

 

 

Gurumurthi, K

159

Rao, P S

171

Yasodha, R

159

 

 

Rathore, J S

241

 

 

 

 

Rathore, V

241