International Research Journal of Biological Sciences ___________________________________ ISSN 2278-3202Vol. 2(9), 69-75, September (2013) Int. Res. J. Biological Sci. International Science Congress Association        
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Assessment of six Mulberry (Morus) Germplasm varieties through Moulting and Bioassay parameters by using Crossbreed Silkworms Bombyx mori L. for Commercial Exploitation in Kolar District, Karnataka, INDIAVeerapura Narayanappa YoganandaMurthy*, Hyadalu Lingappa Ramesh, Gangadharaiah Lokesh, Munirajappa and Bangalore Ramakrishna Dayakar YadavDepartment of Life Sciences, Ganga Kaveri Institute of Science and Management, Rajajinagar, Bangalore-560021, Karnataka, INDIA Department of Sericulture, V.V.Pura College of Science, K.R. Road, Bangalore-560004, Karnataka, INDIA 
Silkworm Division, Central Tasar Research and Training Institute, Central Silk Board, Ranchi-835303, Jharkhand, INDIA Department of Sericulture/Life Sciences, Jnana Bharathi Campus, Bangalore University, Bangalore-560056, Karnataka, INDIA Mulberry Pathology Division, Central Sericultural Research and Training Institute, Central Silk Board, Mysore-570008, Karnataka, INDIAAvailable online at: 
www.isca.in
Received 7th June 2013, revised 16th July 2013, accepted 6th August 2013Abstract  A critical assessment study comprising six mulberry germplasm varieties viz., Tr, Tr12, Tr20, Matigara black, Morus nigra and M was conducted at Bethamangala village in Bangarpet taluk, Kolar district for their leaf quality through economic characters of silkworms. Silkworm rearing experiments were performed using crossbreed (PM x NB) silkworms. Taxa studied differ significantly in respect of moulting and bioassay trials. Moulting test results revealed that, silkworm larvae reared on Tr8 mulberry leaves recorded highest values and lowest values were recorded in silkworm larvae reared on Morus nigra leaves both in I and II moult respectively. Silkworm rearing performance revealed that, silkworms reared on Trmulberry leaves proved better and lower values were observed in silkworms reared on Morus nigra leaves in all the bioassay parameters. From the results, it is proved that, mulberry variety Tr turns out to be a superior in moulting and bioassay tests compared to other varieties studied under the same agro climatic conditions.    Keywords: Mulberry genotypes,evaluation, bombyx mori, cocoon characters, feeding. Introduction Silkworm Bombyx mori L. is an economically important insect used for silkworm rearing and silk production essentially a monophagous insect feeds solely on mulberry leaves Morus spp.) as the sole natural food. Silkworm larval growth and development and cocoon crop yield are mainly influenced by yield and nutritional quality of mulberry (Morus spp.) leaf used as feed1-4. Evaluation of mulberry plants aimed at selecting superior varieties for rearing performance mainly depends on leaf quality. It is a known fact that, around 60% of the total cost of cocoon production goes towards mulberry production alone in sericulture. Hence, in recent years maximum attention has been given for the improvement of mulberry in terms of both quality and quantity. Quality of mulberry varieties fed for silkworms plays a vitol role in the economy of sericulture industry. Among the various factors influencing silkworm growth and cocoon production, leaf quality plays a major role8,9. It is a confirmed fact that, leaf quality differs among mulberry varieties which in turn responsible for the difference in silkworm rearing performances10. Increased production of superior quality mulberry leaves with high nutritive value enhances the chances of good cocoon crop and successful sericulture11,12. In the present experiment an attempt has been made to screen better performing mulberry variety through silkworm rearing experiment in Kolar district is one of the premier and traditional sericulture belt accounts for 40% of the total raw silk production in Karnataka state.   Material and Methods In the present study mulberry varieties like Tr, Tr12, Tr20, Matigara black, Morus nigra and M were used.  mulberry variety is used as a check variety for the comparison purpose. Cuttings of these varieties were procured from CSGRC, Hosur, Tamil Nadu, India and disease free cross breed (PM x NB) egg layings obtained from National Silkworm Seed Project (NSSP), Bangalore, Karnataka were used. Mulberry plants were grown in field at Bethamangala village in Bangarpet taluk  Kolar district of Karnataka state as it is shown in figure 1. Experiment was conducted in RBD method with 4 replications / variety. Two years old plants were used for silkworm rearing from time to time in different season’s viz., summer, rainy and winter and the average values were tabulated in tables. Silkworm Moulting: Moulting test was carried out up to 2ndmoult following standard rearing methods with four replications/variety and 100 larvae/replication. Tender leaves (1st-4th order) on healthy shoots were harvested and fed to young age silkworm larvae up to second moult13. Silkworm rearing was conducted under standard conditions14,15. Daily three 
International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202   Vol. 2(9), 69-75, September (2013)  Int. Res. J. Biological Sci.     International Science Congress Association 
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feedings were given at 7am, 2pm and 10pm from brushing to end of II moult with tender, succulent and nutritious leaves. First appearance of one larva out of moult was considered as commencement of moulting16. Comparative moulting ratio with respect of all the varieties leaves under evaluation was fixed depending on time duration which recorded more than 50% of the larvae under moult. Larval weight was also recorded. Silkworm Rearing: As in figure 2, Silkworm rearing was conducted to test the efficiency of selected mulberry varieties. Rearing experiments were conducted at different seasons (rainy: July-August, winter: Nov-Dec, summer: March-April). For each mulberry variety, one egg laying was reared and four replications were maintained. After III moult, about 100 larvae / replication were maintained. Appropriate cellular rearing techniques were adopted and separate rearing trials were conducted for different varieties17,18. Leaves were harvested during cooler hours of the day and preserved in wet gunny cloth till the feeding time. Larvae were fed three times daily (7am, 2pm, 10pm) with healthy, fresh mulberry leaves. Young age larvae were fed with tender, succulent and nutritious leaves which are known to favour the growth and development of chawki silkworms, while mature and coarse leaves were fed to larvae when they grow till ripening. Cocoons shown in figure 3 were collected on 5th day of mounting and were assessed for commercial cocoon characters. Standard methods employed for the assessment of cocoon quality19.  Statistical Analysis: Data collected on various parameters were tabulated and subjected to critical statistical analysis by adopting ‘Method of Analysis of Variance’ appropriate to the design of the experiment20,21. Figure-1 Map showing location of experimental area: Bethamangala in Kolar District 
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Results and Discussion Silkworm (Bombyx mori L.) larval growth and development is vary depending on the quality and quantity of mulberry leaf used as food source, which in turn indicated by commercial characteristics of cocoon crop22,23,24,25. Several reports are available on the evaluation of mulberry varieties through silkworm rearing performances26-29. It is quite evident that tender, succulent and nutritious leaves are known to favour the good growth and development of young age silkworms whereas progressively mature leaves with less moisture content are required for late age silkworms30. Figure-2 th instar Crossbreed (PM X NB) Silkworms   Figure-3 Crossbreed (PM X NB) Cocoons Silkworm Moulting: Highly significant differences were observed in larval weight and moulting ratio in silkworm larvae fed with selected mulberry varieties as shown in table 1. Silkworms reared on Tr leaves revealed highest single larval weight (4.02mg) and moulting ratio (74.97%) and lowest single larval weight (2.74mg) and moulting ratio (66.04%) was observed in Morus nigraleaves in I moult. In II moult, high single larval weight (23.90mg) and moulting ratio (83.70%) was observed in silkworms reared on Tr leaves and lowest single larval weight (15.94mg) and moulting ratio (70.04%) was recorded in Morus nigra leaves. Degree and uniformity of moulting varies with mulberry leaf quality that favours the higher moulting ratio, ensures better growth rate and silkworm larval weight. It was reported that, higher proteins and lower sugars in S41 variety supported high larval weight and moulting ratio31,32. Lower larval weight and moulting ratio in Mysore local variety were associated with lower leaf moisture content and moisture retention ability. It was recorded that, 89.16% and 92.82% moulting ratio in PM x NB and NB18 x NB races respectively with S54 mulberry genotype leaves33. When CB and BV larvae fed on Tr10 and MR mulberry varieties leaves have showed higher moulting ratio and larval weight34. Mulberry genotypes S30, S36 and Vishwa were superior over M genotype in moulting ratio and larval weight35.  Silkworm Rearing: Results revealed that, highly significant differences were observed in bioassay parameters and cocoon characters of silkworms reared on mulberry varieties as tabulated in table 2. Weight of ten 5th instar larvae (36.95g) was found significantly higher in silkworms reared on Tr leaves and lower larval weight (27.86g) was recorded in silkworms reared on Morus nigra leaves. Single cocoon weight (1.71g) was higher in cocoons obtained from silkworms reared on Trleaves. However, significantly lower cocoon weight was (1.38g) registered in silkworms reared on Morus nigra leaves. Shell weight and percentage were significantly high in cocoons obtained from silkworms rearing on Tr leaves (0.35g and 21.18% respectively) while the cocoons obtained by rearing silkworms on Morus nigra leaves recorded lower shell weight and shell weight percentage (0.23g and 16.24%) respectively. Filament length and reelability percentage was significantly high in cocoons obtained from silkworms reared on Tr leaves (926.81mts, 77.50% respectively). Lower filament length and lower reelability was recorded in cocoons harvested from silkworms reared on Morus nigra leaves (786.32mts, 59.69%). Significantly lower renditta (6.68) was recorded in the cocoons procured from silkworms reared on Tr leaves, while higher renditta (7.86) was found in the cocoons recovered from silkworms reared on Morus nigra leaves. Finer denier was recorded in cocoons procured from silkworms reared on Trleaves (2.16) while cocoons procured from silkworms reared on Morus nigra leaves (2.82) produced coarser denier. Effective rate of rearing was significantly higher in silkworms reared on Tr leaves (87.04%) and lower effective rate of rearing was recorded in silkworms reared on Morus nigra leaves (79.25%). Berhampore variety was better than Kosen and Mandalaya with regard to effective rate of rearing and cocoon weight22. Mulberry variety Catteneo was found best in silkworm rearing trials compared to Burmose, Tsukasakhu and Local mulberry varieties36. Mulberry variety S showed distinct varietal effect 
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on all economic characters37. Mulberry variety S54 recorded higher values in rearing parameters and coon yield38. S41 and S54mulberry varieties supported good larval growth and recorded high cocoon weight compare to cocoons obtained from S36while Mysore local and Kanva registered lowest cocoon weight39.  Mulberry variety S54 was found to be superior followed by S41 and Kanva for feeding silkworms40,41. Feeding of Ichinose leaves to silkworms resulted in higher cocoon characters compared to other two varieties42. Silkworm larvae fed with Kokuso27 variety leaves revealed best results among other varieties studied43. Mulberry variety S30 showed better performance than S36, S41 and K for commercial characters of bivoltine cocoons44. Mulberry variety S36 was found superior in silkworm rearing trials and recorded highest cocoon yield, ERR and shell percentage45. Among the seven mulberry cultivars tested for rearing parameters by using bivoltine silkworms and observed that, C763 performed best followed by S1630 and C1730 mulberry varieties46. Silkworm breeds (PM x NB) and (NB) performed well when both the races larvae reared on M mulberry variety leaves47,48. Mulberry variety RFS175 was found superior in silkworm rearing trials with maximum cocoon production49. It was reported that, mulberry variety S1635 was found superior in silkworm rearing trials with good commercial characters of cocoons when compared to all other varieties examined50,51,52. Mulberry varieties Tr10 and MR were found superior as they secured maximum scores for silkworm races CB (PM x NB) and BV (NB) in rearing tests53. According to many scientists rearing performance of silkworm races differed significantly when they are subjected to same conditions, some of them performed better and poor performance by some races54,55,56. Present study also confirms the same as S1708 mulberry variety gives better results in pre-cocoon and post-cocoon characters when compared to other varieties tested. Among eight mulberry varieties i.e. S, S146, S1635, AR12, AR14, TR10, BR and Kevaluated for nutritional potential by silkworm rearing experiments, silkworms reared on BR leaves showed higher larval weight and cocoon parameters compared to other varieties57. Out of the ten mulberry varieties like Tr, Tr12, Tr20, 1708, MSMatigara black, C, C10, Morus nigra and Mvarieties screened for their nutritional superiority through bioassay experiments, bivoltine (NB) silkworm reared on 1708 and Tr leaves recorded significantly high moulting ratio, high larval weight and commercial cocoon parameters compared to other varieties58. Three silkworm races namely EC, EJ and EJ were evaluated for their growth and productivity by feeding them with S36 mulberry leaves and reported that, EJ silkworms showed a higher and consistent growth rates compared to other silkworm races for all the developmental stages59. Rearing performance of bivoltine silkworm hybrid NB x SH reared during spring season revealed good performance in respect of cocoon parameters such as single cocoon weight (1.81g) and shell weight (0.384g)60. Cocoon weight and shell weight are the most important characters evaluated for productivity61. Shell weight percentage indicates the amount of raw silk can be reeled from the given quantity of fresh cocoons and shell weight percentage varies according to age and breed of silkworm. Total silk filament length is ranging from 600m-1500m out of which only 80% is reelable62. In the present study, silk filament length of cocoons recovered from silkworms reared on different mulberry varieties falls within this range and cocoons recovered from silkworms reared on Tr mulberry variety leaves produced longest filaments length with lowest denier.  Table-1 Crossbreed silkworms (PM X NB) moulting performance on different mulberry varieties Mulberry varieties I Moult II Moult 
Single larval  weight (mg) Moulting ratio (%) Single larval  weight (mg) Moulting ratio (%) 
Tr 4.02 74.97 23.90 83.70 
Tr12 3.95 74.60 23.51 82.81 
Tr20 3.72 73.63 23.30 81.88 
Matigara black 3.10 71.80 21.19 77.85 
Morus nigra 2.74 66.04 15.94 70.04 
 3.66 72.81 22.59 80.78 
CD @ 5% 0.01 0.03 0.03 0.03 
 
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Table-2 Crossbreed silkworms (PM X NB) bioassay performance on different mulberry varieties Mulberry varieties Weight of 10
 
th instar larvae(g) Single cocoon weight (g)
 
Single shell weight (g)
 
Shell weight percentage (%) Filament length (mts) Reelability percentage (%) Renditta
 
Denier
 
ERR (%) 
Tr 36.95 1.71 0.35 21.18 926.81 77.50 6.68 2.16 87.04
Tr
12
 34.95 1.64 0.35 20.25 879.16 76.25 6.96 2.29 83.68
Tr
20
 33.38 1.63 0.32 19.81 860.43 75.00 7.25 2.38 86.40
Matigara black 29.10 1.45 0.26 17.71 804.73 62.50 7.66 2.71 82.59 
Morus nigra 27.86 1.38 0.23 16.24 786.32 59.69 7.86 2.82 79.25 
M
5
 32.16 1.54 0.30 19.59 830.86 97.50 7.28 2.50 84.56
CD @ 5% 0.36 0.05 0.01 0.37 19.01 7.06 0.09 0.02 0.14 
Conclusion It is evident from the present studies that, moulting ratio, silkworm larval weight and rearing performance of crossbreed silkworms is significantly high when silkworms fed with Trmulberry variety leaves. Commercial cocoon characters like finer denier, lower renditta were recorded high in cocoons procured from silkworms reared on Tr leaves compared to other mulberry varieties studied. Next to Tr variety, Tr12 and Tr20 varieties were proved promising followed by check variety . These varieties leaves supported good growth and development of silkworm larvae,that reflected in better commercial cocoon characteristic features. Mulberry varieties Matigara black and Morus nigra occupied the last place in bioassay results. From the results, it is concluded that, Trmulberry germplasm variety turns out to be a superior in moulting and bioassay tests compared to other varieties studied under the same agro climatic conditions. Such genotype may be recommended for more trials at field level by the farmers and could be exploited for commercial purpose under Kolar area for sustainable growth and development of sericulture industry.    References 1.Adolkar V.V., Raina S.K. and Kimbu D.M., Evaluation of various mulberry Morus spp. (Moraceae) cultivars for the rearing of the bivoltine hybrid race Shaanshi BV-333 of the silkworm Bombyx mori (Lepidoptera: Bombycidae), Int. J. Trop. Insect Sci., 27(1), 6-14 (2007)2.Bongale U.D. and Chaluvachari., Evaluation of eight mulberry germplasm varieties by leaf biochemical and bio-assay moulting studies, Sericologia,35, 83-94 (1995)3.Nagaraju J., Application of genetic principles in improving silk production, Current Science, 83(4), 409-415 (2002) 4.Purohit K.M. and PavanKumar T., Influence of various agronomical practices in India on the leaf quality in mulberry A review, Sericologia, 36(1), 27-39 (1996) 5.Bongale U.D., Chaluvachari., Mallikarjunappa R.S., Narahari Rao B.V., Anantharaman M.N. and Dandin S.B., Leaf nutritive quality associated with maturity levels in fourteen important varieties of mulberry (Morus spp.), Sericologia, 37(1), 71-81 (1997)6.Chaudhary P.C. and Giridhar K., Mulberry cultivation. In: Appropriate Sericulture Techniques, Ed. by Jolly MS, ICTRTS, Chapter-2, Mysore, India, 17-34 (1987)7.Das B.C. and Sikdar A.K., Evaluation of some improved strains of mulberry by feeding experiment, Indian J. Seric.,9(1), 26-30 (1970)8.Matsumara, On the functional difference of digestive amylase of different strains of the silkworm Bombyx moriL. Bull. Seric. Exp. Sta.Japan,13, 513-519 (1951) 9.Bose P.C., Evaluation of mulberry leaf quality by chemical analysis, In: Genetic resources of mulberry and utilization,Ed. by Sengupta, K. & Dandin, S.B. CSR&TI, Mysore, India, 189-190 (1989)10.Arug H., Principles of sericulture. Oxford and IBH publishing Co.Pvt.Ltd, 66 Janpath New Delhi, India, 376 (1994)11.Krishnamurthy K.C., Mulberry cultivation in the tropics, Asian women in tropical sericulture, United Nations Economic and Social Commission for Asia and the Pacific, ST/ESCAP/247, 12-13 (1983)12.Ravikumar C., Western ghat as a bivoltine region prospects, challenges and strategies for its development, Indian Silk, 26(9), 39-54 (1988)13.Benchamin K.V. and Nagaraj C.S., Silkworm rearing techniques, In: Appropriate sericulture techniques, Ed. by M.S. Jolly, Chapter-4, ICTRTS, Mysore, India, 63-106 (1987) 14.Krishnaswami S., New technology of silkworm rearing, Bulletin No.2, Central Silk Board, Bangalore, India, 1-23 (1986b)15.Krishnaswami S., Improved method of rearing young age (chawki) silkworms, Bulletin No.2, Central Silk Board, Bangalore, India, 1-24 (1990)
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41.Tayade D.S., Jawale M.D. and Unchegaonkar P.K., Evaluation of some improved strains of mulberry by feeding experiments under Maratawada conditions, Indian J. Seric.,27(1), 92-94 (1988)42.Dar H.U., Singh T.P. and Das B.C., Evaluation of mulberry varieties by feeding to Bombyx mori., Indian J. Seric., 27(1), 16-22 (1988)43.Fotadar R.K., Ahsan M.M., Dhar K.L. and Bhakuni B.S., Evaluation and utilization of genetic variability in mulberry, Indian J. Seric., 28(2), 150-158 (1989)44.Sathyanarayana Raju Ch., Pallavi S.N., Reddy M.M., Suryanarayana N., Singhal B.K. and Sengupta K., Evaluation of four new mulberry varieties through silkworm rearing under irrigated condition, Indian J. Seric., 29(2), 240-247 (1990)45.Saratchandra B., Rajanna L., Philomena K.L., Paramesha C., Ramesh S.P., Jayappa T. and Sabitha M.G., An evaluation of elite mulberry varieties for yield and quality through bio-assay, Sericologia, 32(1), 127-134 (1992) 46.Das B.K., Saker J., Das C., Das N.K. and Sen S.K., Seasonal effects on the relative performance of five bivoltine breeds of silkworm Bombyx mori L., Uttar Pradesh Journal of Zoology, 15(2), 91-96 (1995) 47.Changalarayappa and Chinnaswamy K.P., Influence of mulberry varietal feeding sequences on growth and development of silkworm (Bombyx mori L.) breeds, Proc. Natl. Semi. Tropical Seric., Bangalore, India, 204-205 (1999)48.Vage M.N. and Ashoka J., Performance of fifth instar silkworm of PM x NB on tender shoot feeding for larval and cocoon yield traits, Proc.Natl.Semi.Tropical Seri.,Bangalore, India, 97-99 (1999)49.Rachotaiah M., Narahari Rao B.V., Prabhuraj D.K. and Dandin S.B., Status of biochemical constituents in four mulberry varieties grown under rainfed condition and their effect on silkworm, Bombyx mori L. Natl. Conf. Stra. Seri. Res. Devpt., Mysore, India, 41 (2000)50.Chakravorty R. and Borgohain P.N., Identification of an improved mulberry variety suitable for north eastern part of India, Indian J. Seric., 39(1), 74-75 (2000) 51.Rahman M.S., Doss S.G., Vijayan K. and Roy B.N., Performance of the mulberry variety S1635 under three systems of planting in West Bengal, Indian J. Seric., 38(2),165-167 (1999) 52.Santosha Gowda V. Patil., Evaluation of promising genotype S1635 under irrigated conditions, Indian Silk, 41(2),7-9 (2002)53.Sujathamma P., Dandin S.B. and Savithri G., Quality evaluation of mulberry (Morus spp.) genotypes through bioassay under Royalaseema conditions of Andhra Pradesh, Indian J. Seric., 40(1), 27-34 (2001)54.Fonseca T.C., Almeida J.E. and Fonseca A.S., Effect of mulberry selection on silkworm feeding. Sericologia, 30(4),475-477 (1990)55.Giridhar K. and Reddy S.N., Effective rate of rearing in bivoltine silkworm, Bombyx mori breeds on different mulberry, Morus sp. Varieties, Indian J. Seric., 30(1), 88-90 (1991) 56.Seidavi A., Evaluation of the genetic potential of six native strains of silkworm Bombyx mori L. African Journal of Agricultural Research, 6(20), 4816-4823 (2011)57.Gangwar S.K., Impact of varietal feeding of eight mulberry varieties on Bombyx mori L. Agriculture and Biology Journal of North America, 1(3), 350-354 (2010)58.Yogananda Murthy V.N., Ramesh H.L., Munirajappa and Dayakar Yadav B.R., Nutritional quality assessment of ten mulberry (Morus) germplasm varieties through moulting Test, silkworm rearing techniques and economical characters of bivoltine silkworms (Bombyx mori L) for commercial exploitation, Int. Res. J. Natural Sci.1(2), 11-22 (2013)59.Ogunleye R.F. and Johnson O., Determination and comparison of performance and production properties in three strains of silkworm bombyx mori (l) fed with a variety of mulberry plant, Int. Res. J. Biotechnol., 3(5), 76-80 (2012)60.Singh Amardev., Ahmed Farooq and Bandey Shamin Ahmed, Spring rearing performance by feeding temperate mulberry variety on bivoltine hybrid NB4D2 x SH6 of silkworm, Bombyx mori L., ISCA Journal of Biological Sciences., 1(2), 69-72 (2012)61.Gaviria D.A.E., Aguilar H.J., Serrano and Alegria A.H., DNA fingerprinting using AFLP markers to search for makers associated with yield attributes in the silkworm Bombyx mori ., J. Insect Sci.6, 1-10 (2006)62.FAO, Silk reeling and testing manual, Agricultural Services, Bulletin No. 136 Rome, Italy, 65-75 (1999)
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