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.     

Development of new sources of cytoplasmic male-sterile lines in rice

Two male-sterile lines, KalashreeA and PadminiA, with a Miz.21 cytoplasm source were developed through indica/indica hybridization followed by repeated backcrossing with their respective recurrent male parents (Kalashree and Padmini) up to the BC₆ generation. These two cytoplasmic male-sterile lines are suitable for use in the development of hybrids for lowland situations owing to their intermediate to semi-tall stature, late flowering duration, good grain quality and easy fertility restoration ability.     

Feed consumption, nutrient utilization and serum metabolite profile of captive blackbucks (Antelope cervicapra) fed diets varying in crude protein content

A feeding trial was conducted to determine the optimum level of crude protein (CP) in the diet of captive blackbuck (Antelope cervicapra) in which feed consumption and nutrient utilization are maximal. Fifteen blackbucks (BW 25-34 kg) were distributed into three groups of five each in an experiment of 75-days duration including a digestion trial of 5-day collection period. All the animals were offered 200 g of concentrates and fresh maize fodder ad libitum. The overall CP content of the three respective diets was 6.9%, 10.4% and 12.7%. Blood samples were collected on the last day of the experiment. Intake and digestibility of CP increased (p < 0.01) with the increased level of CP in the diet. Feed consumption and nutrient intake were not significantly different among the groups. However, digestibilities of most of the nutrients were higher in the 10.4% CP diet than in the 6.9% CP diet. The endogenous loss of nitrogen was similar among the groups. Based on the endogenous losses, minimum N requirement was calculated to be 776 mg/kg BW(0.75) /day, and to meet this requirement, diet must contain at least 8.27% CP. Serum urea nitrogen concentration increased (p < 0.01) with increased level of dietary CP. Serum level of glutamate oxaloacetate transaminase and glutamate pyruvate transaminase was higher (p < 0.05) in the group fed 6.9% CP diet. Animals in the group fed low protein diet also lost body mass during the experimental period. It was concluded that a diet containing 10.4% CP was optimum for maximizing nutrient utilization without any adverse effect on voluntary feed consumption and serum metabolite profile of blackbucks.     

Immunogene expression in head kidney and spleen of common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.) following thermal stress and challenge with Gram-negative bacterium, <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

To evaluate the effect of thermal and microbial stress on the immune response of common carp (Cyprinus carpio L.), relative mRNA expression level of pro-inflammatory cytokines [tumor necrosis factor alpha (TNF-α) and interleukin (IL)-1β] and other genes related to immune or stress response [inducible nitric oxide synthase (iNOS), heat shock protein 70 (Hsp70), superoxide dismutase one (SOD1), and glucocorticoid receptor (GR)] was measured by quantitative PCR (qPCR). In addition, total protein and total immunoglobulin level in blood plasma of experimental common carp was also assayed. All the above parameters were estimated 24 h post-challenge with Gram-negative bacterium, Aeromonas hydrophila. Common carp (54.89?±?6.90 g) were initially exposed to 20 °C (control group) and 30 °C (thermal stress group) water temperature for 30 days, followed by experimental challenge with 2.29?×?10⁸ colony forming unit/mL (CFU/mL; LD₅₀ dose) of A. hydrophila. Exposure of fish to thermal stress and subsequently challenge with A. hydrophila significantly (P?<?0.05) increases the IL-1β mRNA expression in head kidney and spleen of common carp by ~?39.94 and ~?4.11-fold, respectively. However, TNF-α mRNA expression in spleen decreased ~?5.63-fold in control fish challenged with A. hydrophila. Thermal stress and challenge with bacterium suppresses the iNOS and GR mRNA expression in spleen of common carp. Moreover, significant (P?<?0.05) increase in total protein content of blood plasma (~?43 mg/g) was evident in fish exposed to thermal stress and challenged with A. hydrophila. In conclusion, our study highlights the importance of elevated temperature stress and microbial infection in differential regulation of expression of several immunogenes in common carp.     

Status of floor vegetation under some monoculture and mixculture plantations in North India

A study conducted to assess the floor vegetation shows the presence of 17 species underPopulus deltoides in comparison to 28, 29 and 35 underAlbizia lebbeck, Dalbergia sissoo and mix-culture plantations, respectively. Further, the biomass and density of floor vegetation is considerably reduced underP. deltoides compared to other plantations. In addition, the indices for species richness, evenness, dominance and diversity are also reduced underP. deltoides showing some inhibitory mechanism being operative under these plantations. Based on available literature, allelopathy is speculated to be responsible for this reduction.     

Morphological and molecular variability among Indian isolates of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> causing banded leaf and sheath blight in maize

Rhizoctonia solani, a devastating soil borne fungus inciting banded leaf and sheath blight (BLSB) disease is a constraint in maize production and improvement program. Rhizoctonia isolates collected from seven diverse maize cropping zones of India were examined for morphological and molecular variability. All the tested isolates caused symptoms of BLSB on maize and were also cross infective on rice and sugarcane hosts, but showed significant variability in hyphal diameter, mean hyphal cell size, weight, size and distribution of scleorotia, culture pigmentation, incubation period, pathogenicity and expression of symptoms. Neighbour joining cluster analysis placed the 62 isolates of R. solani into four major groups, A, B, C and D. Group A was more diverse and included isolates of diverse agro-ecological zones. The cluster analysis corresponded well with principle component analysis. Pathogenicity testing of R. solani isolates on maize genotype (CM 501) revealed highly variable virulence pattern of the pathogen population suggesting its high evolutionary potential, and hence adaptability to diverse geographical regions. The study reveals a strong evidence of inherent potential of the R. solani isolates to survive in diverse ecological zones and its probable spread to other maize cultivars across India. Sequence comparisons of the internal transcribed sequence-ribosomal DNA region of 62 isolates did not reveal much diversity among the isolates. Majority of the isolates (n?=?61) clustered together with anastomosis group (AG) AG1-IA used as reference strain in the phylogram, distinct from AG1-IB, AG2–2IIIB and Waitea circinata used as reference strains. BLSB isolates representing distinct geographical locations shared identical sequences indicating long-distance dispersal of the pathogen. The study confirms that the genetic flexibility of the pathogen allows for its adaptation to variable ecological niches and long-distance introduction of new genotypes into the region. The study emphasizes that epidemiological studies may complement the molecular studies.     

Moisture-dependent physical properties of jatropha fruit

The physical properties of fruit are important in designing and fabricating equipment and structures for handling, transporting, processing and storage, and also for assessing quality. The study was conducted to investigate some physical properties of jatropha fruit at various moisture levels. The average length, width, thickness and 1000 mass were 29.31 mm, 22.18 mm, 21.36 mm and 1522.10 g, respectively, at moisture content of 7.97% d.b. The geometric mean diameter increased from 24.03 to 24.70 mm and the sphericity varied between 0.82 and 0.83 as moisture content increased from 7.97% to 23.33% d.b., respectively. In the same moisture range, the bulk and true densities decreased from 278 to 253 and 546 to 435 kg m^?3, respectively, whereas the corresponding porosity also decreased from 49.08% to 41.84%. As the moisture content increased from 7.97% to 23.33% d.b., crushing strength was decreased from 275 to 79 N, whereas the angle of repose and surface areas were found to increase from 36.41° to 41.72° and 1815.73 to 1917.59 mm², respectively. The static coefficient of friction of jatropha fruit increased linearly against the surfaces of three structural materials, namely plywood (47.81%), mild steel (62.88%) and aluminium (34.82%) as the moisture content increased from 7.97% to 23.33% d.b.     

Low Temperature Effects during Seed Filling on Chickpea Genotypes (Cicer arietinum L.): Probing Mechanisms affecting Seed Reserves and Yield

Chickpea is sensitive to cold conditions (<15 °C), particularly at its reproductive phase and consequently it experiences significant decrease in the seed yield. The information about the effects of cold stress on chickpea during the seed filling phase is lacking. Moreover, the underlying metabolic reasons associated with the low temperature injury are largely unknown in the crop. Hence, the present study was undertaken with the objectives: (i) to find out the possible mechanisms leading to low temperature damage during the seed filling and (ii) to investigate the relative response of the microcarpa (Desi) and the macrocarpa (Kabuli) chickpea types along with elucidation of the possible mechanisms governing the differential cold sensitivity at this stage. At the time of initiation of the seed filling (pod size ∼1 cm), a set of plants growing under warm conditions of the glasshouse (temperature: 17/28 ± 2 °C as average night and day temperature) was subjected to cold conditions of the field (2.3/11.7 ± 2 °C as average night and day temperature), while another set was maintained under warm conditions (control). The chilling conditions resulted in the increase in electrolyte leakage, the loss of chlorophyll, the decrease in sucrose content and the reduction in water status in leaves, which occurred to a greater extent in the macrocarpa type than in the microcarpa type. The total plant weight decreased to the same level in both the chickpea types, whereas the rate and duration of the seed filling, seed size, seed weight, pods per plant and harvest index decreased greatly in the macrocarpa type. The stressed seeds of both the chickpea types experienced marked reduction in the accumulation of starch, proteins, fats, crude fibre, protein fractions (albumins, globulins, prolamins and glutelins) with a larger decrease in the macrocarpa type. The accumulation of sucrose and the activity levels of the enzymes like starch synthase, sucrose synthase and invertase decreased significantly in the seeds because of the chilling, indicating impairment in sucrose import. Minerals such as calcium, phosphorous and iron as well as several amino acids (phenylalanine, tyrosine, threonine, tryptophan, valine and histidine) were lowered significantly in the stressed seeds. These components were limited to a higher extent in the macrocarpa type indicating higher cold sensitivity of this type.     

Exogenous Application of Abscisic Acid Improves Cold Tolerance in Chickpea (Cicer arietinum L.)

Chickpea suffers cold stress (<10 °C) damage especially during reproductive phase resulting in the abortion of flowers and pods, poor pod set, and reduction in seed yield and seed quality. One of the ways in modifying cold tolerance involves exogenous treatment of the plants with chemicals having established role in cold tolerance. In the present study, the chickpea plants growing under optimum temperature conditions (28/12 °C, as average maximum and minimum temperature) were subjected to cold conditions of the field (10–12/2–4 °C; day/night as average maximum and minimum temperature) at the bud stage. Prior to exposure, these plants were treated exogenously with 10 μm abscisic acid (ABA) and thereafter again after 1 week of exposure. The stress injury measured in terms of increase in electrolyte leakage, decrease in 2,3,5-triphenyl tetrazolium chloride reduction %, relative leaf water content and chlorophyll content was observed to be significantly mitigated in ABA-applied plants. A greater pollen viability, pollen germination, flower retention and pod set were noticed in ABA-treated plants compared with stressed plants. The seed yield showed considerable improvement in the plants treated with ABA relative to the stressed plants that was attributed to the increase in seed weight, greater number of single seeded pods and reduction in number of infertile pods. The oxidative damage measured as thiobarbituric acid-reactive substances was lesser in ABA-treated plants that was associated with greater activities of superoxide dismutase, catalase, ascorbate peroxidase, ascorbic acid, glutathione and proline in these plants. It was concluded that cold stress effects were partly overcome by ABA treatment because of the improvement in water status of the leaves as well as the reduction in oxidative damage.