Potential,constraints and applications of in vitro methods in improving grain legumes

Grain legumes, the important constituents of sustainability‐based cropping systems and energy‐limited vegetarian diets have long been the subject of scientific research. Tremendous technological strides were made in the so‐called orphan crops, in terms of both varietal improvement and generation of basic information. Despite recalcitrancy and high genotype dependency, in vitro culture techniques such as organogenesis, in vitro mutagenesis, embryo rescue and in vitro gene transfer have been deployed for improvement of several grain legumes and these played an important role in introgression of desirable genes from related and distant species and creation of additional genetic variability. Stable and reproducible regeneration protocols resulted in the development of genetically modified chickpea, pigeon pea, cowpea, mungbean, etc., while embryo rescue was deployed successfully for recovery of interspecific recombinants, a few of them exploited for the development of commercial cultivars. Nevertheless, doubled haploidy witnessed limited success and protoplast regeneration and in vitro mutagenesis remained of academic interest. The present review focuses on the progress, achievements, constraints and perspectives of using in vitro technology in grain legume improvement.     

Cryopreservation of silver barb Puntius gonionotus (Bleeker) spermatozoa: effect of extender composition,cryoprotective agents and freezing rate on their postthawing fertilization ability

The effects of extender composition, cryoprotectant concentration and freezing and thawing on the fertilization efficiency of cryopreserved spermatozoa of Puntius gonionotus were evaluated. Computer‐aided motility analysis of semen was conducted to check the suitability of spermatozoa for cryopreservation after mixing with different extenders and cryoprotective agents (CPAs). Extender‐4 with an osmolality 260 mOsmol kg⁻¹and pH 7.6 was used for the cryopreservation study. Among the CPAs, dimethyl sulphoxide (DMSO) was least toxic and more than 60% fertilization was achieved when used at 1.4 M at 0 °C for 10 and 30 min, whereas the toxicity of all CPAs to spermatozoa was evident when tested at 30 °C. Semen frozen at −16 °C min⁻¹ with 1.4 M DMSO showed 70% fertilization, which was significantly higher (P<0.05) than other freezing rates. Samples thawed at 35 °C water showed a fertilization rate comparable with that of fresh semen. Computer‐assisted semen analysis of fresh and frozen semen after thawing showed variations in different types of motility in spermatozoa and in their class. There was no significant difference in motility before or after cryopreservation; however, significant differences could be observed in the average path velocity (VAP), straight line velocity (VSL) and curve linear velocity (VCL). Semen of silver barb could be cryopreserved with extender‐4 by addition of 1.4 M DMSO to a final cryopreservation medium (MED 2) cooled at a rate of −16 °C min⁻¹, stored in liquid nitrogen (−196 °C) and utilized after thawing at 35±2 °C.     

Effect of dietary Curcuma longa on enzymatic and immunological profiles of rohu,Labeo rohita (Ham.), infected with Aeromonas hydrophila

The aim of this study was to evaluate the dietary dosages of turmeric that enhance immune response and disease resistance against the opportunistic pathogens Aeromonas hydrophila in Labeo rohita fingerlings. Hence, four different dosages of turmeric at 0.1, 0.5, 1.0 and 5.0 g kg^?1 feed were given to the fingerlings of L. rohita for 60 days at 4% body weight. At every 20‐day interval, different biochemical, haematological, enzymatic and immunological parameters of fish were evaluated. After 60 days, fish were challenged with A. hydrophila. The mortality (%) was recorded on the tenth day post challenge. Most of the immune parameters including lysozyme activity, superoxide anion production and serum bactericidal activity were significantly (P<0.05) higher on 60 days of feeding of 1.0 g of turmeric per kg of feed. Challenge study indicated 100% and 89% survivability in the group of fish fed with 5.0 and 1.0 g of turmeric per kg of feed respectively. Feeding of turmeric might have maintained long‐term protection in fish by elevating the nonspecific immune system such as Nitroblue tetrazolium (NBT), lysozyme and serum bactericidal activity. The result showed that turmeric at a dose of 1.0 g kg^?1 feed for 60 days provided the greatest protection to pathogen challenge.     

Influence of crop establishment methods on yield,economics and water productivity of rice cultivars under upland and lowland production ecologies of Eastern Indo-Gangetic Plains

A field study on assessment of crop establishment methods on yield, economics and water productivity of rice cultivars under upland and lowland production ecologies was conducted during wet seasons (June–November) of 2012 and 2013 in Eastern Indo-Gangetic Plains of India. The experiment was laid-out in a split-plot design (SPD) and replicated four times. The main-plot treatments included three crop establishment methods, viz. dry direct-seeded rice (DSR), system of rice intensification (SRI) and puddled transplanted rice (PTR). In sub-plots, five rice cultivars of different groups like aromatic (Improved Pusa Basmati 1 and Pusa Sugandh 5), inbreds (PNR 381 and Pusa 834) and hybrid (Arize 6444) were taken for their evaluations. These two sets of treatments were laid-out simultaneously in two production ecologies, upland and lowland during both years. In general, lowland ecology was found favourable for rice growth and yield and resulted in 13.2% higher grain yield as compared to upland ecology. Rice grown with SRI method produced 19.4 and 7.0% higher grain yield in 2012 and 20.6 and 7.1% higher in 2013, over DSR and PTR. However, PTR yielded 13.1 and 14.5% higher grain over DSR during 2012 and 2013, respectively. On an average, Arize 6444 produced 26.4, 26.9, 28.9 and 54.7% higher grain yield as compared to PS 5, P 834, PNR 381 and IPB1, respectively. Further, the interaction of production ecologies × crop establishment methods revealed that, in upland ecology, SRI recorded significantly higher grain yield as compared to PTR and DSR, but in lowland, grain yield resulting from SRI was similar to the yield obtained with PTR and significantly higher than DSR. The latter two methods (PTR and DSR) yielded alike in lowland ecology in both study years. The production ecologies × crop establishment methods × cultivars interaction on grain yield showed that the growing of Arize 6444 cultivar using SRI method in upland ecology resulted in the higher grain yield (8.87 t/ha). But the cost of production was also highest in SRI followed by PTR and DSR across production ecologies and cultivars. Cultivation of hybrid (Arize 6444) involved higher cost of production than all other cultivars. Irrespective of crop establishment methods and cultivars, gross returns, net returns and B:C ratio were significantly higher in lowland compared to upland ecology. Owing to higher grain yield, SRI method fetched significantly higher gross returns and net returns over PTR and DSR. Average increase in net return with Arize 6444 was 68.8, 41.0, 37.7 and 33.1% over IPB 1, PNR 381, P 834 and PS 5, respectively. There was a saving of 30.7% water in SRI and 19.9% in DSR over PTR under upland ecology. Similarly in lowland ecology, water saving of 30.2% was observed in SRI and 21.2% in DSR over PTR. Due to higher yield and saving on water, SRI returned significantly higher total water productivity (TWP) (5.9 kg/ha-mm) as compared to DSR (3.5 kg/ha-mm) and PTR (3.6 kg/ha-mm) under upland ecology. In lowland ecology, also SRI (6.2 kg/ha-mm) resulted in higher TWP as compared to other two methods. However, DSR gave significantly higher TWP as compared to PTR. Among cultivars, hybrid Arize 6444 recorded the highest TWP in both upland and lowland production ecologies across crop establishment methods. Hence, growing of hybrid Arize 6444 with SRI method can enhance rice productivity and water-use efficiency in lowland and upland production ecologies of Eastern Indo-Gangetic Plains and in other similar regions.     

Integrated effect of population and weed management regimes on weed dynamics,performance, and productivity of basmati rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Weeds caused serious problem on yield reduction of basmati rice worldwide. Losses caused by weeds varied from one country to another, depending on the presence of dominant weeds and the control methods practiced by farmers; therefore, suitable plant population and weed management practices should be adopted. Keeping these in mind, a field experiment was carried out during kharif seasons of 2009 and 2010 at crop Research Centre of SVPUA&T, Meerut, India comprising 4 planting geometries, viz. 20, 30, 40, and 50 hills m^?2 as main plot factor, and 5 weed management practices (Butachlor @ 1.0 kg ha^?1, Butachlor @ 1.0 kg ha^?1 fb (followed by) one hand weeding, Butachlor @ 1.0 kg ha^?1 fb Almix @ 4 g ha^?1, two hand weedings and weedy check) in a split plot design with 3 replications. Experimental results revealed that plant population of 50 hills m^?2 proved superior over that of 20 hills m^?2 in respect of weed density, weed dry weight, number of tillers m^?2, yield attributes, grain, straw, and biological yields. The maximum grain yield (29.00 and 31.00 q ha^?1) and straw yield (51.30 and 52.50 q ha^?1) were recorded in 50 hills m^?2 followed by 40 hills m^?2 during 2009 and 2010, respectively. In respect of nitrogen, phosphorus, and potassium removal, a reverse trend was observed: the highest in 20 hills m^?2 followed by 30, 40, and 50 hills m^?2. As far as the weed management practices are concerned, both chemical and mechanical methods of weed control were found superior over weedy check. The lowest weed density, dry weight, and highest weed control efficiency, maximum length of panicle^?1, number of panicle (m²), and 1000-grain weight and grain yield of 30.40 and 32.60 q ha^?1 were recorded with two hand weedings which was at par with Butachlor @ 1.0 kg ha^?1 fb one hand weeding over rest of the weed management practices.     

Alteration in important quality traits and antioxidant activities in Brassica oleracea with Ogura cybrid cytoplasm

In vegetable brassicas, cytoplasmic male sterility (CMS) system is the most preferred mechanism for hybrid seed production and Ogura cybrid cytoplasm is the only source used widely. Effects of the alien cybrid cytoplasm on important quality traits need to be understood for their effective use in breeding programme. In analysing 38 Ogura cybrid cytoplasm‐based cauliflower CMS lines, it was revealed that introgression of Ogura cytoplasm resulted in significant alteration in important quality traits. Cupric reducing antioxidant activity and ferric reducing ability of plasma values were increased up to 5–10 times in different genotypes. Among 38 analysed CMS lines, concentration of plant pigments such as anthocyanin, total chlorophylls and ascorbic acid was reduced in 21, 17 and 32 genotypes, respectively. However, the concentration of total carotenoids and β‐carotene was elevated in most of the CMS lines and total carotenoid was increased up to 20 times in the CMS line, Ogu13‐85‐2A. The results indicate the role of nuclear–cytoplasmic interaction and mitochondrial genome in determining concentration of different quality traits.     

Extension of shelf life of tuberose flowers using a combination of gamma irradiation and generally regarded as safe (GRAS) preservatives and assessment of antimicrobial potency of senesced flowers

Tuberose cut flowers, available as loose flowers, were treated with gamma (γ) irradiation and generally regarded as safe (GRAS) preservative solutions for extension of shelf life. The flowers were packaged in low-density polyethylene bags, heat sealed and stored at 23 ± 2°C, 80% relative humidity (RH) and 4 ± 1°C, 40% RH, respectively. The flowers stored at these two temperature regimes were subjected to sensory evaluation and biochemical analyses. From these assessments, the longest shelf life of tuberose flowers was found to be 8 days at 23 ± 2°C, 80% RH (compared to 4 days for control) and 24 days at 4 ± 1°C, 40% RH (compared to 8 days for control) using combination treatment of low dose γ-irradiation (0.02 kGy) and preservative solutions (4% sucrose and 0.02% CaCl₂). Ethanolic extract of tuberose flowers of the most shelf stable set (stored at 4 ± 1°C), i.e. at the end of 24 days, showed antimicrobial potency against the common skin pathogen Staphylococcus aureus (ATCC 25923 and MDR strains), suggesting utility of the senesced tuberose flowers for therapeutic applications. This preservation technique would promote export of tuberose flowers by extension of their shelf lives and allow utilization of these flowers, post senescence.     

Comparisons of energy balance and evapotranspiration between flooded and aerobic rice fields in the Philippines

The seasonal and annual variability of sensible heat flux (H), latent heat flux (LE), evapotranspiration (ET), crop coefficient (K_c) and crop water productivity (WP_ET) were investigated under two different rice environments, flooded and aerobic soil conditions, using the eddy covariance (EC) technique during 2008-2009 cropping periods. Since we had only one EC system for monitoring two rice environments, we had to move the system from one location to the other every week. In total, we had to gap-fill an average of 50-60% of the missing weekly data as well as those values rejected by the quality control tests in each rice field in all four cropping seasons. Although the EC method provides a direct measurement of LE, which is the energy used for ET, we needed to correct the values of H and LE to close the energy balance using the Bowen ratio closure method before we used LE to estimate ET. On average, the energy balance closure before correction was 0.72 ± 0.06 and it increased to 0.99 ± 0.01 after correction. The G in both flooded and aerobic fields was very low. Likewise, the energy involved in miscellaneous processes such as photosynthesis, respiration and heat storage in the rice canopy was not taken into consideration.Average for four cropping seasons, flooded rice fields had 19% more LE than aerobic fields whereas aerobic rice fields had 45% more H than flooded fields. This resulted in a lower Bowen ratio in flooded fields (0.14 ± 0.03) than in aerobic fields (0.24 ± 0.01). For our study sites, evapotranspiration was primarily controlled by net radiation. The aerobic rice fields had lower growing season ET rates (3.81 ± 0.21 mm d⁻¹) than the flooded rice fields (4.29 ± 0.23 mm d⁻¹), most probably due to the absence of ponded water and lower leaf area index of aerobic rice. Likewise, the crop coefficient, K_c, of aerobic rice was significantly lower than that of flooded rice. For aerobic rice, K_c values were 0.95 ± 0.01 for the vegetative stage, 1.00 ± 0.01 for the reproductive stage, 0.97 ± 0.04 for the ripening stage and 0.88 ± 0.03 for the fallow period, whereas, for flooded rice, K_c values were 1.04 ± 0.04 for the vegetative stage, 1.11 ± 0.05 for the reproductive stage, 1.04 ± 0.05 for the ripening stage and 0.93 ± 0.06 for the fallow period. The average annual ET was 1301 mm for aerobic rice and 1440 mm for flooded rice. This corresponds to about 11% lower total evapotranspiration in aerobic fields than in flooded fields. However, the crop water productivity (WP_ET) of aerobic rice (0.42 ± 0.03 g grain kg⁻¹ water) was significantly lower than that of flooded rice (1.26 ± 0.26 g grain kg⁻¹ water) because the grain yields of aerobic rice were very low since they were subjected to water stress.The results of this investigation showed significant differences in energy balance and evapotranspiration between flooded and aerobic rice ecosystems. Aerobic rice is one of the promising water-saving technologies being developed to lower the water requirements of the rice crop to address the issues of water scarcity. This information should be taken into consideration in evaluating alternative water-saving technologies for environmentally sustainable rice production systems.     

Earthworm biomass response to soil management in semi-arid tropical Alfisol agroecosystems

Earthworms recorded during 1989–1993 across 15 soil management treatments, comprising three different tillagexthree organic amendments (bare, farmyard manure, and rice straw) and six perennial ley treatments, belonged to two endogeic species, Octochaetona phillotti (Michaelsen) and Lampito mauritii Kinberg, while in a nearby undisturbed natural revegetation area three species were found, including the above two and Octonochaeta rosea (Stephenson). The earthworm biomass showed significant temporal and spatial variations and was higher during the post monsoon period than in the early rainy season. No worm biomass was recorded during the dry season. In the tillage and organic amendment treatments, the biomass was drastically reduced from September 1989 to September 1991 after the application of carbofuran and some herbicides, and was significantly reduced during these two years compared to that of 1992. The maximum monthly earthworm biomass ranged between 2.5 and 17.9 g m^-2 across the treatments and increased several-fold in the nearby natural revegetation area (75.9 g m^-2). It significantly increased in perennial ley treatments compared to annual treatments with organic amendments. Thus the earthworm biomass varied significantly (P<0.01) across the 15 treatments, indicating discernible effects of soil management.Visiting Scientist (under the Rockefeller Foundation Environmental Research Fellowship in International Agriculture)     

Methane emission from two Indian soils planted with different rice cultivars

In a greenhouse study, methane emissions were measured from two diverse Indian rice-growing soils planted to five rice cultivars under similar water regimes, fertilizer applications and environmental conditions. Significant variations were observed in methane emitted from soils growing different cultivars. Total methane emission varied between 8.04 and 20.92gm^–2 from IARI soil (Inceptisol) and between 1.47 and 10.91gm^–2 from Raipur soil (Vertisol) planted to rice. In all the cultivars, emissions from IARI soil were higher than from Raipur soil. The first methane flux peak was noticed during the reproductive phase and the second peak coincided with the grain-ripening stage of the rice cultivars. Received: July 7, 1996