Journal of Environment and Sociobiology

Pijush Mallick ¹, Sayantan Ghosh ¹, Shruti Chattaraj ¹, Samir Ranjan Sikdar ¹

Affiliations
1 Division of Plant Biology, Bose Institute, P-1/12, C.I.T., Scheme VII M, Kolkata-700 054, West Bengal, IN

Abstract

Mulberry (Morus alba L. cv S1635) is a perennial crop plant in Indian sericulture and used for silkworm Bombyx mori L. feeding. Qualitative and quantitative improvement of this crop is highly essential in respect to better and quality silk production and hence through breeding approaches the development of new lines is important. Protoplasts are the good source of any plant to study about their genetics and cellular mechanisms for breeding. Using cell wall degradating enzymes mulberry protoplasts from mesophyll tissue were isolated. Several combinations of cellulose, macerozyme and driselase enzymes mixture were tested in different incubation conditions. A combination of Cellulase R10-1%, Macerozyme R10-1% and Driselase-0.25% in 0.6M mannitol showed comparatively better yield (3.8 × 106 / gm tissue) after 3-4 hrs shaking with 100 rpm at 28°C.

Keywords

Mulberry Plant, Mesophyll Protoplast, Cell Wall Degradating Enzymes.

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Bisweswar Bera ¹, Priyanka Samanta ¹, Samir Ranjan Sikdar ², Pijush Mallick ²

Affiliations
1 Department of Biotechnology, School of Biotechnology & Biosciences, Brainware University, 398, Ramkrishnapur Road, Barasat, North 24 Parganas, Kolkata-700125, IN
2 Division of Plant Biology, Bose Institute, P-1/12, CIT Rd, Scheme VIIM, Kankurgachi, Kolkata-700054, IN

Abstract

Towards establishment of biodiversity character via analysis of relative DNA content of wild edible mushroom, Termitomyces species is used here for the first time. Dye propidium iodide (PI), an intercalating fluorescent molecule with molecular mass of 668.4 Da is used for staining of Termitomyces protoplasts to analyse the cellular DNA content using flow cytometer. PI binds the cellular nucleic acids at 535 nm excitation and 617 nm emission range. The protoplasts were isolated from liquid MYG grown vegetative tissues of edible mushroom strain Termitomyces heimii. The isolated protoplasts were purified with 0.6M manitol (MW 3350) followed by washing in PBS buffer. Protoplast pellet was suspended in PI (l mg/ml) hypotonic solution and final volume made up with sterile water. Incubation was done at 37°C for 30 min followed by analyzing using flow cytometer. Results showed that staining depends upon the protoplast purity, size and viability. The vegetative tissue showed a heterogeneous protoplast population in terms of sizes which is ultimately reflected on the relative amount of DNA content. The heterogeneous protoplast population in the suspension through cytometry score results the unequal amount of DNA content in every cell meaning unequal size, age and viability. It is established that PI can be used for staining of mushroom protoplast in biodiversity and genetic research.

Keywords

Edible mushroom, Protoplast, Propidium iodide, Flow cytometry, DNA content.

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Sayantan Ghosh ¹, Samir Ranjan Sikdar ²

Affiliations
1 School of Biotechnology and Biosciences, Department of biotechnology, 398, Ramkrishnapur Road, Brainware University, Barasat, West Bengal: 700 125, IN
2 Bose Institute, Division of Plant Biology, P-1/12, C. I. T. Road, Scheme VII M, Kolkata-700 054, West Bengal, IN

Abstract

Population abundance of mustard aphid Lipaphis erysimi (Kalt.) was studied upon the treatment of aqueous extract from eight cultivars of Indian mustard Brassica juncea L. from different agronomical regions of India. A wild crucifer Rorippa indica, known for its aphid tolerant nature, also was studied, side by side. Treatment of R. indica extract on the isolated population of mustard aphids showed a significant suppressing effect on aphid population. Cultivar Gujarat Mustard 3 was appeared to be best host for the aphid, and cv. Ashirwad was seen to be most tolerant to L. erysimi as on the treatment of water extract from Gujarat Mustard 3.Nymphal growth of aphid was not affected and a regular growth pattern was observed here. But aqueous extract treatment from cv. Ashirwad significantly controlled aphid population on non-host Nicotiana tabacam.

Keywords

Mustard Aphids, Brassica Juncea, Lipaphis Erysimi, Rorippa Indica, Aqueous Extract.

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Neelanjana Bag ¹, Bijay Pal ¹, Samir Ranjan Sikdar ², Pijush Mallick ¹

Affiliations
1 Department of Biotechnology, School of Biotechnology & Biosciences, Brainware University, 398, Ramkrishnapur Road, Barasat, North 24 Parganas, Kolkata-700125, IN
2 Division of Plant Biology, Bose Institute, P-1/12, CIT Rd, Scheme VIIM, Kankurgachi, Kolkata-700054, IN

Abstract

A total of twelve somatic hybrid strains, Pleurotus florida x Lentinus squorrosulus (pfls), were characterized through PCR based RAPD analysis. Sixteen random decemer primers yielded a total of 136 fragments with 97.79 % polymorphism with an amplicon range of 200 bp - 3000 bp in size. RAPD-08 showed optimum amplification and generated 12 scorable bands (250 bp -1800 bp) where RAPD-07 amplified least and generated 5 scorable bands (400 bp - 1300 bp). PIC ranged from 0.424 (SRS-15) to 0.499 (SRS-16). Based on Jaccard’s proximity matrix the highest genetic distance was found in between parent L. squarrosulus and hybrid pfls1q which means they are genetically most distantly related. Among the hybrid pfls1k and pfls1p showed maximum genetic distance while the most closely related hybrids were pfls1i and pfls1l. The parent L. squarrosulus was most genetically distant individual than the other parent P. florida.

Keywords

Hybrid Mushroom, Genetic Diversity, Polymorphism, RAPD.

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Shruti Chattaraj ¹, Samir Ranjan Sikdar ¹

Affiliations
1 Division of Plant Biology, Bose Institute, P-1/12, C.I.T. Scheme VII M, Kolkata-700 054, IN

Abstract

The fatty acid profile from fruit bodies of four somatic hybrid mushroom (AbCi) strains and their parent strains Calocybe indica var. APK2 and Agaricus bisporus were studied using GC-MS. The study showed that Palmitic acid and Stearic acid were the most abundant saturated fatty acids (SFA) and this ranged from 15.0 - 30.18% in the hybrids and parent strains. Among mono-unsaturated fatty acids (MUFA), Oleic acid was found abundantly in hybrids AbCi 6, AbCi 7 and AbCi 8 compared to any of the parents. Poly-unsaturated fatty acids (PUFA) dominated among all the hybrids than their parents and maximum PUFA was found in AbCi 7 (64.6 %).In hybrids (AbCi 7- 59.95 % and AbCi 9- 60.4 %) and parents Linoleic acid dominated. Overall the unsaturated fatty acids were at higher concentrations than saturated fatty acids in hybrids than in parent strains.

Keywords

Fatty acid, Hybrid mushroom, Edible mushroom, GC-MS

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