Volume 4 Number 1 2010

CONTENTS AND ABSTRACTS

Amalendu Tikader, Chandrakant Kamble (India) Seri-biodiversity with Reference to Host Plants in India (pp 1-11)

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Invited Review: India harbours a wide range of biodiversity due to its diverse climatic and cultural practices. There are five types of natural silkworms, which feed on host plants of mulberry, arjun, asan, sal, Quercus, som, soalu, castor, kesseru, bar kesseru, tapioca and payam. Unlike mulberry silkworm, the wild silkworms are polyphagous and feed on several plants. Mulberry is the leading silk and 1120 host plants are being maintained with 13 species at the Central Sericultural Germplasm Resources Centre, Hosur. Tropical tasar host plant is represented by 13 species, distributed in 20 states and 124 genetic resources of host plant are being maintained in the ex situ gene bank at the Central Tasar Research and Training Institute, Ranchi. In temperate tasar a total of 10 species of oak are growing in the eastern part of the country and 6 species in the Western Himalayas for silkworm rearing. About 14 morphotypes ofsom and 10 morphotypes of soalu are maintained for muga silkworm. The primary food plant of eri silkworm is castor and 41 accessions are maintained. Although mulberry dominates silk variety in the global silk market, other non-mulberry silks also have importance in the domestic market. Besides this primary and secondary food plants have many multipurpose uses other than for silk. The availability, biodiversity, progress of work in different aspects to utilize and conserve plant genetic resources is reviewed and discussed.

Ehab AbouKheir, Trichy Ganesh Prasad, Madavalam Sreeman Sheshshayee, Makarla Udayakumar (India) Dual Stable Isotopes Carbon (¹³C) and Oxygen (¹⁸O) – An Approach to Identify Desirable Cotton Lines (pp 12-18)

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Original Research Paper: Enhancement of crop productivity under drought stress can be achieved by maximizing soil water capture by a deep or dense root system and by effective water use by increasing intrinsic ability to fix more carbon per unit water transpired (water use efficiency, WUE). Although the importance of these characters for improving productivity under drought is well known, precise quantification of these traits under field conditions in a large number of genotypes is the major limitation in breeding crop plants for these traits. In recent years, several reports showed the relevance of using stable isotope composition of carbon and oxygen in biomass as a rapid and precise technique to determine the genetic variability in WUE and transpiration rate, respectively in plants. In this study, genetic variability in both carbon and oxygen stable isotopes was examined among 15 upland cotton (Gossypium hirsutum) lines and significant genetic variability was noticed. We confirmed the relationship between Δ¹³C with WUE and Δ¹⁸O with transpiration rate among cotton lines. Therefore, measurement of stable isotopes can be used as a surrogate for time-averaged measurement of WUE and transpiration rate. We demonstrate that cotton lines with low Δ¹³C and high Δ¹⁸O accumulated higher biomass due to high WUE coupled with high photosynthetic capacity while maintaining relatively high transpiration. We illustrate the efficiency of selection of cotton lines based on dual stable isotopes (carbon and oxygen) to identify higher biomass.

Esmail Bakhshandeh, Rahman Ghadiryan, Behnam Kamkar (Iran) A Rapid and Non-Destructive Method to Determine the Leaflet, Trifoliate and Total Leaf Area of Soybean (pp 19-23)

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Original Research Paper: The importance of rapid, non-destructive, and accurate measurements of leaf area for agronomic and physiological studies is well known. Several mathematical formulas have been derived for estimating leaf areas for numerous crops, but there is little information available for soybean (Glycine max L.). This study aimed to develop prediction equations for estimating leaflet, trifoliate and total leaf areas using maximum length, L (cm), width, W (cm), length and width product, LW (cm) and green leaf dry matter, DM (g) of soybean leaves. For this purpose, an experiment was conducted using three cultivars of soybean (‘Dpx’, ‘Sahar’ and ‘Williams’), in 2009-2010, in the Faculty of Agronomy, Gorgan, Iran. During the growing season, leaves of randomly selected soybean plants were collected. Leaf area was measured with a digital leaf area meter, related dry matter also was weighed, leaf dimensions were determined with a ruler, too. Statistical analyses of soybean leaf areas were divided into three levels: leaflet, trifoliate and total leaf area. At each level, the predictive abilities of three regression equations (linear, power and binomial) were compared, with different independent variables for each equation. Our data indicate, however, that considerable savings of time, with little loss of predictive ability, could be possible by measuring only W or LW in each instance. In general, these analyses indicated that a single regression equation could be used at each level. Our findings revealed that pooled-based models (without respect to cultivar) are reliable for estimating leaflet, trifoliate and total leaf area. Researchers can use these models readily and without any inconvenience to save time and costs, especially where there is a lack of related equipment to measure leaf area.

Mohd. Idrees, M. Masroor A. Khan, Tariq Aftab, M. Naeem (India) Synergistic Effects of Gibberellic Acid and Triacontanol on Growth, Physiology, Enzyme Activities and Essential Oil Content of Coriandrum sativum L. (pp 24-29)

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Original Research Paper: Two well known growth regulators, triacontanol (TRIA) and gibberellic acid (GA₃), were applied on economically as well as medicinally important plant coriander (Coriandrum sativum L.) and their effects were noted in the present study. The plants were sprayed with either TRIA or different combinations of TRIA and GA₃ at 30 days after sowing. Among the treatments, a foliar spray of 10^-6 T + 10^-6 G significantly promoted the values for most of the growth (shoot and root lengths, fresh and dry weights), physiological and biochemical (total chlorophyll and carotenoid, nitrate reductase activity, carbonic anhydrase activity, leaf nitrogen and leaf phosphorus content except leaf potassium content) parameters, including the essential oil content and yield characteristics (number of umbels per plant, number of fruits per umbel, 100-seed weight and seed yield). On the basis of data obtained from the present work, it may be concluded that a combined spray of TRIA and GA₃ (10^-6 T + 10^-6 G) on coriander plant is highly effective for productivity with increased essential oil content.

M. Naeem, M. Masroor A. Khan, Moinuddin, Minu Singh (India) Calcium Chloride Stimulates Crop Yield, Photosynthetic Efficiency, Enzyme Activities and Nutraceuticals of Coffee Senna (Senna occidentalis L.) under Calcium-Deficient Soil (pp 30-37)

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Original Research Paper: Plant biological yield appears to be comparatively low in calcium-deficient soil of Aligarh, western Uttar Pradesh, India. Ca deficiency poses a serious yield and quality limitation for several crops, including medicinal herbs, in this region of India. In view of the importance of coffee senna as a medicinal legume, it was hypothesized that calcium application through soil could enhance crop productivity, photosynthetic efficiency, enzymatic activities and nutraceuticals. Plants were grown in pots containing soil supplied with five levels of calcium, viz. 0, 40, 80, 120 and 160 mg Ca kg^-1 soil (Ca₀, Ca₁, Ca₂, Ca₃ and Ca₄, respectively) applied as calcium chloride (CaCl₂). The performance of the crop was assessed in terms of various growth, physiological, biochemical, yield and quality attributes at 120, 270, 300 and 330 days after sowing. Calcium application effectively increased most of the parameters studied. Of the five calcium levels, Ca₃ stimulated most of the attributes studied at the three growth stages (120, 270 and 300 DAS). In fact, Ca₃ increased seed-yield and seed-protein content by 27.6 and 10.6%, respectively, compared to control plants.

Swarnendu Chandra, Soumya Chatterjee, Krishnendu Acharya (India) Biological Control of Fruit Rot and Die Back Disease of Capsicum annuum with Pseudomonas aeruginosa WS-1 (pp 38-42)

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Original Research Paper: Colletotrichum capsici is the causal agent of fruit rot and die back disease, the most important diseases of chilli worldwide. A rhizospheric isolate Pseudomonas aeruginosa WS-1 successfully showed both in vitro (10⁷ cells/ml)and in vivo (3 × 10⁸ cfu/ml)antagonistic activity against the pathogen C. capsici. On dual plate culture, the antagonist effectively suppressed the growth of the fungal pathogen. Light microscopy of mycelium from the interacting region showed hyphal shriveling, hyphal deformities, tip swelling, short branching and granulation of cytoplasm resulting in lysis of hyphae. The antifungal activity of this isolate is correlated with the production of siderophores and HCN. The strain also produces mycolytic enzyme-like proteases and chitinases, which are well known to lyse the cell walls of the fungal pathogen. The efficacy of P. aeruginosa WS-1 application under greenhouse conditions was comparable to that of fungicide (carbendazim) to manage the disease. This result suggests that P. aeruginosa WS-1 has the potential to be used as a biopesticide for effective management of fruit rot and die back disease of chilli in India.

S. Sree Lekha, J. Sree Kumar, Santha V. Pillai (India) Assessing Genetic Diversity of Indian Cassava: A Comparison of Old and New Collections Using Microsatellite Markers (pp 43-52)

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Original Research Paper: In this study the variability and diversity in cassava (Manihot esculenta Crantz) at the molecular level was assessed using SSR (simple sequence repeat) markers. 60 accessions of old collections and 50 accessions of new collections from different parts of South India were compared to study their genetic diversity. 36 sets of SSR markers, available in the public domain and known to generate polymorphic bands, were used for amplification. There were significant differences in genetic diversity between old and new collections. The similarity index among the old varieties ranged from 80 to 100% and from 68 to 98% in the new varieties. 34 loci or 34.34% of the loci were polymorphic in old collections and 59 or 93.65% of the loci were polymorphic in the new collection. The Shannon’s Information Index for old varieties was 0.2778 and for new varieties it was 0.4967 for older ones. Wright’s mean fixation index was higher than the mean value expected (5%) indicating an excess of homozygotic individuals in old collections. The presence of new recombination events by random and natural processes of mutation and recombination might have resulted in high genetic diversity in new collections.

Gaurav Aggarwal, Amarjit Kaur Nath, Chhaya Sharma, Satya Vrat Bhardwaj, S.K. Chakarovarty (India) Characterization of Prunus armeniaca L. Germplasm using RAPDs (pp 53-56)

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Original Research Paper: The present study characterizes apricot germplasm through the use of RAPD markers. Twelve apricot cultivars (‘Chuli’, ‘Suffaida’, ‘Shakarpara’, ‘Kaisha’, ‘Nari’, ‘Royal’, ‘Charmagaz’, ‘Shipley Early’, ‘New Castle’, ‘Tilton’, ‘Turkey’ and ‘Moorpork’) were selected. RAPD analysis was carried out using 18 random decamer primers, 16 of which showed polymorphism. A total of 79 bands were amplified, out of which 56 were polymorphic and 10 were specific RAPD markers. Total polymorphism was 70.89%. The similarity value ranged from 0.59 to 0.86. A low similarity value was obtained between ‘Shakarpara’ and ‘Chuli’ cultivars and high between ‘Moorpork’ and ‘Suffaida’ cultivars. On the basis of a similarity matrix, a dendrogram was constructed by the UPGMA method. RAPD analysis is useful in a certification scheme for releasing certified plant material. It could also be useful for assessing genetic variation among different apricot accessions.

Shabir H. Wani, Ajaz A. Lone (India), Jaime A. Teixeira da Silva (Japan), Satbir S. Gosal (India) Effects of NaCl Stress on Callus Induction and Plant Regeneration from Mature Seeds of Rice (Oryza sativa L.) (pp 57-61)

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Original Research Paper: Two commercial indica rice varieties, PAU 201 and PR 116, exhibited appreciable callus induction in vitro but the capacity to induce and regenerate callus decreased under salinity stress in both when exposed to sodium chloride (NaCl) at 90 mM. Callus was induced on semi-solid MS medium supplemented with 2.5 mg/l 2,4-dichlorophenoxyacetic acid + 0.5 mg/l kinetin + 560 mg/l proline + 30 g/l sucrose + 8 g/l agar. Callus developed shoots on MS medium supplemented with 2.0 mg/l benzylamino purine + 0.5 mg/l kinetin + 0.5 mg/l α-naphthalene acetic acid + 30 g/l sucrose + 8 g/l agar. The results indicated the need to optimize the protocol for callus induction, maintenance and regeneration before selection process for tolerance to salinity.

S. D. Shruthi, Y. L. Ramachandra, S. Padmalatha Rai, Prashant Kumar Jha (India) Pharmacognostic Evaluation of the Leaves of Kirganelia reticulata Baill. (Euphorbiaceae) (pp 62-65)

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Original Research Paper: The leaves of Kirganelia reticulata Baill. (Euphorbiaceae) are known to have many uses in Indian ethnomedicine. Establishment of a pharmacognostic profile of the leaves will assist in standardization for quality, purity and sample identification. Evaluation of the fresh, powdered and anatomical sections of the leaves were carried out to determine the macro- and micromorphological characters, and quantitative, qualitative and phytochemical profiles. The results of the study could be useful in setting some diagnostic indices for the identification and preparation of a monograph of the plant.

Ahmed M. Eed, Solipuram Amarender Reddy, Kankanala Malla Reddy (India), Jaime A. Teixeira da Silva (Japan), Punnam Veera Reddy, Hameedunnisa Beghum, Pedda Y. Venkatsubbaiah (India) Effect of Antibiotics and Fungicides on the in Vitro Production of Citrus limonia Osbeck Nodal Segment and Shoot Tip Explants (pp 66-70)

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Original Research Paper: Among the several rootstocks, Rangpur lime (Citrus limonia Osbeck) is commercially employed in India, particularly in Andhra Pradesh state, for the propagation as a rootstock for sweet orange cultivar ‘Sathgudi’ on account of its disease tolerance, drought resistance and overall better performance over other cultivars. The in vitro propagation of plants is always under threat of microbial and fungal contamination either from the environment or from the explants. To overcome the surface contamination of explants, several experiments were conducted to evaluate the efficacy of different antibiotics and fungicides. The treatments consist of benomyl (0.01%), bavistin (0.1%) and antibiotics viz., ampicillin, streptomycin and tetracycline at 150, 200 and 250 mg/L. The growth medium used was Murashige and Skoog (MS) supplemented with 3% sucrose and 0.5 mg/L benzylaminopurine (BAP). Benomyl (100 mg/L) and streptomycin (250 mg/L), when added to MS medium after surface sterilization, resulted in 93.33 and 100% bacterial and fungal-free explants, respectively.

Gunnaiah Raghavendra, Gollarahalli Kenchappa Kumaraswamy, Boregowda Ramya, Hakkare Swamidatta Sandesh, Kalenahalli Narasimhamurthy Yogendra, Nanjappa Deepak, Paramanahally Hanumanthegowda Ramanjini Gowda (India) Direct Multiple Shoot Regeneration of indica Rice (Oryza sativa) Var. ‘Rasi’ (pp 71-73)

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Short Communication: Tissue culture of indica rice cultivars is difficult compared to japonica cultivars as they are recalcitrant to in vitro culture. In vitro culture of rice through an intervening callus stage not only poses the problem of somaclonal variation but also takes a very long time to regenerate. Rapid regeneration of indica rice variety ‘Rasi’ was carried out from the shoot basal portion, which produced multiple shoots on Murashige and Skoog (MS) medium containing thidiazuron (TDZ) and 6-benzyl amino purine. MS medium with 4 mg/L TDZ produced 11 shoots/explant with 93.3% regeneration efficiency. Shoots rooted on MS medium with 0.5 mg/L α-naphthalene acetic acid and were transferred to hardening medium containing equal volumes of sterilized sand and vermicompost within 45 days of culture. In vitro grown plants were uniform and no somaclonal variation was observed.

Rajagopal Chitra, E. Vadivel (India) In Vitro Organogenesis in Phyllanthus amarus Schum. and Thonn. (pp 74-76)

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Short Communication: A procedure is outlined for organogenesis of Phyllanthus amarus using nodal segment and shoot tips. Multiple shoot formation was induced from the explants on Murashige and Skoog medium (MS) supplemented with benzyl amino purine (BAP) and kinetin (Kin). MS + BAP (4.0 mgl^-1) + Kin (4 mgl^-1) recorded maximum multiple shoot from shoot and nodal segments. Among the explants, nodal segments gave higher number of shoots (5.94 shoots/explant) than shoot tip (5.22 shoots/explant). The shoot tip derived plants (95%) showed higher survival percentage than nodal segments (88%). Addition of GA₃ to the medium enhanced the shoot length. On sub-culturing the multiple shoots in MS medium supplemented with BAP (2.0 mgl^-1) and GA₃ (0.5 mgl^-1) had resulted in a 7-fold increase in shoots. Rooting (87%) of the shoots was best achieved in ½ MS + IBA (0.5 mgl^-1) + IAA (0.5 mgl^-1). The plantlets were hardened in a mixture of sand and vermiculite (1:1) and successfully established in natural soil, where they grew and matured normally.

I. N. Bheema Lingeswara Reddy, D. Srinivas Reddy, M. Lakshmi Narasu, S. Sivaramakrishnan (India) Development and Evaluation of SCAR Marker for Blast Resistance in Finger Millet (Eleusine coracana (L.) Gaertn) (pp 77-80)

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Research Note: Blast disease is caused by the fungus Pyricularia grisea and is a widespread disease in India and other countries affecting different aerial parts of the plant at all stages of its growth starting from seedling to grain formation. Specific DNA technology, such as the use of SCAR (sequence characterized amplified region) markers, can be used to differentiate between resistant and susceptible species of finger millet (Eleusine coracana (L.) Gaertn). SCAR markers were created by sequencing a single RAPD (random amplified polymorphic DNA) band and designing primers to amplify the band of specific size. PCR analysis was done using two F₂ bulks (one resistant and another susceptible) and two resistant F₃ bulk populations. RAPD primer OPA20 amplified bands of 250 and 750 bp in resistant bulks and absent in susceptible bulks. The fragments were cloned and converted into SCAR markers. Two SCAR primer pairs were designed. Testing was done with 3 resistant cultivars and one susceptible cultivar; single unique bands were correctly amplified for resistant cultivars by SCAR primer pairs. The availability of DNA markers linked to genes for resistance to this disease would accelerate breeding programs, particularly when other traits are also being evaluated.

Ashok K. Agarwal, Govind S. Rajwar (India) Macrophytes of Tehri Dam Reservoir, Garhwal Himalaya, India (pp 81-85)

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Research Note: One hundred and seventy six species of macrophytes (154 herb, 14 shrub and 8 tree species) belonging to 58 families of angiosperms were recorded in the present study on macrophytes of Tehri Dam reservoir in Garhwal Himalaya during 2003-2005. All species were herbs. The family Asteraceae was the most dominant among the macrophytes consisting of 25 species. The second largest family was Papilionaceae with 16 species followed by Poaceae with 14 species. The largest genus was Ageratum with 4 species. Cyperus, Euphorbia, Sida, Crotalaria and Ranunculus had 3 species each. Twenty genera consisted of 2 species each and all other genera had one species each. Comparatively, fewer macrophytes appeared in July and August, showing thinner growth, while maximum growth was recorded in December and January.