Relationship between endometritis and oxidative stress in the follicular fluid and luteal function in the buffalo

In this study, alteration in the follicular fluid composition and luteal function was investigated in the buffalo with endometritis. Genitalia were classified into cytological and purulent endometritis on the basis of polymorphonuclear cell cut off while non‐endometritis served as control (n = 10/group). In the follicular phase, the number of surface follicles was counted, diameter of the largest follicle was measured and the follicular fluid was assayed for total protein, cholesterol, malondialdehyde (MDA), total antioxidant capacity (TAC), oestradiol (E₂) and progesterone (P₄). The P₄ content of corpus luteum during mid‐luteal phase was estimated by radioimmunoassay. Ovaries from the follicular phase of oestrous cycle showed no significant difference in the total number of surface follicles, size of the largest follicle and volume of follicular fluid in the buffaloes with and without endometritis (p > .05). However, the antral fluid of the largest follicle from the genitalia of buffalo with cytological and purulent endometritis showed a significant decrease in the concentration of total protein, cholesterol, TAC and E₂ and a significant increase in the concentration of MDA and P₄ (p < .05). The results indicated that there is an association between endometritis and decreased ovarian function.     

Detection and localization of regucalcin in spermatozoa of water buffalo (Bubalus bubalis): A calcium‐regulating multifunctional protein

Regucalcin (RGN) is a calcium‐regulating, anti‐apoptotic, antioxidative and antiproliferative multifunctional protein predominantly seen in liver and kidney. All these functions are very crucial during spermatogenesis and sperm maturation process until fertilization of the ovum. Although many studies have reported the wide distribution of regucalcin in the male reproductive tract of the rat, human and bovine, its presence in spermatozoa is yet to be demonstrated wherein calcium has a pivotal role in the transport, capacitation, acrosomal reaction and further fusion with ova. Here, we detected the expression of regucalcin mRNA and protein in buffalo spermatozoa using real‐time PCR and Western blot, respectively. The study detected two new regucalcin isoforms of 44 kDa and 48 kDa size along with the reported 34‐kDa, 28‐kDa and 24‐kDa isoforms, wherein the 34‐kDa isoform was found to be membrane associated in spermatozoa. Further, immunocytochemistry study localized the regucalcin protein in the acrosomal region of the caudal and ejaculated buffalo spermatozoa while it was detected in both cytoplasm and acrosomal region of testicular spermatozoa. This discovery of RGN in spermatozoa and localization in the acrosomal region will help to focus researchers to see its role in calcium‐related functions like capacitation, acrosomal reaction and membrane fusion. Overall, regucalcin may be a new fertility marker in buffalo and can be utilized for infertility treatments.     

Antisperm antibodies in repeat‐breeding cows: Frequency,detection and validation of threshold levels employing sperm immobilization,sperm agglutination and immunoperoxidase assay

Antisperm antibodies have been found in repeat‐breeding(RB) cows, and those causing agglutination and/or immobilization of sperm are considered to be closely related to unexplained infertility. However, a standard protocol for identifying antisperm antibodies (ASA) in cattle is not validated. Therefore, an investigation was undertaken to evaluate sperm immobilization (SIT), sperm agglutination (SAT) and immunoperoxidase (IPT)assays for detection of ASA in serum and their respective threshold levels for confirmation. Animals (heifers, normally breeding, repeat‐breeding and pregnant animals) that were free from IBR, brucellosis and uterine infections (screened by clinical examination) were included in the study. Sperm agglutinating, sperm immobilizing and antisperm antibodies evaluated by respective assay were significantly higher (p < .05) in RB cows compared to other groups. The SIT assay was able to identify 61% of RB caused by ASA, more than those employing SAT and IPT. Furthermore, a dilution rate of 1:5 and 1:80 (confirms 59.0 and 57.0% RB+ve)were sufficient to diagnose ASA by SAT and IPT, respectively. Results indicate the presence of __12.6% clumped spermatozoa and __ 2.6%(cut‐off value) peroxidase‐positive spermatozoa at 1:5 and 1:80 dilutions diagnosed with SAT and IPT, respectively, may be considered as repeaters arising out of ASA. Furthermore, study also showed the presence of lower incidence of ASA positivity in other groups of animals (heifer<normal breeder<pregnant animals) compared to repeaters. Study results show that although IPT is more specific and accurate but SAT and SIT are comparatively simple and cost‐effective assays suitable for detecting ASA under field conditions and thus can be recommended for screening of repeaters.     

Conventional and contemporary approaches for drought tolerance rice breeding: Progress and prospects

Drought is becoming a major threat to rice farming across the globe owing to the depletion of water tables in rice-growing belts. Drought affects rice plants at multiple stages, causing damage at morphological and physio-biochemical levels, leading to severe losses that exceed losses from all other stresses. The amalgamation of conventional breeding methods with modern molecular biology tools and biometrical methods could help accelerate the genetic gain for drought tolerance in rice. Many drought-tolerance traits with genetic determinants have been identified and exploited for tolerance rice variety breeding. The integration of genome-wide association study and genomic selection tools with speed breeding shortened the breeding cycle and aided in rapid improvement of genetic gain. In this review, we emphasized the progress made through classical breeding as well as the limitations and usefulness of current genomic methods in improving drought tolerance. We briefly addressed methods for identifying genetic determinants for drought tolerance and deploying them through genomics-assisted breeding programmes to develop high-yielding drought-tolerant rice cultivars.     

Evaluating smallholder farmers’ demand for rice variety attributes in Nepal

Despite the release of many modern varieties of rice (Oryza sativa), farmers in Nepal have been cultivating relatively few rice varieties. The objective of this study is to estimate farmers’ demand for rice variety attributes and to identify the factors that influence their decision regarding varietal choice. Yield increase is considered jointly with other attributes, such as cooking and eating quality, drought tolerance, and maturity period. Choice experiments were conducted and results showed that there was significant demand from farming households for yield increase, improvement in cooking and eating quality, tolerant to drought, and short maturing varieties. The choice data were analyzed using a multinomial and random parameter logit model. From a policy perspective, results showed that for broader adoption of crop varieties, variety development programs should give priority to aforementioned attributes rather than solely on increased crop productivity.     

Genetic diversity and population structure of Indian soybean [<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merr.] revealed by simple sequence repeat markers

Genetic diversity in 90 Indian soybean cultivars was assessed using 45 SSR markers distributed on 20 soybean chromosomes. Forty-five SSR markers generated 232 alleles with an average of five alleles/locus. The observed frequencies of the 232 alleles ranged from 0.01 to 0.94 with an average of 0.19. The polymorphic information content (PIC) value of the SSR markers varied from 0.10 to 0.83 with an average of 0.61 and about 71% markers have a PIC value of >0.5. In this study, 54 rare alleles including 19 genotype specific alleles were also identified. The observed hetrozygosity for SSR markers ranged from 0 to 0.11 with a mean of 0.10. Cluster analysis grouped the 90 soybean cultivars into three major clusters and principal coordinates analysis (PCoA) results were similar to those of the cluster analysis. A combination of eight SSR markers successfully differentiated all 90 soybean cultivars. The population structure analysis distributed the 90 soybean genotypes into two populations with mean alpha (α) value of 0.1873. In AMOVA analysis, proportion of variation within population was high (88%), whereas only 12% occurred among populations. In cluster and structure analyses, most of the genotypes with similar pedigree were grouped together. Soybean cultivars DS228, MACS-13, LSb-1, Hardee, Improved Pelican, and Pusa-24 were the six most genetically distinct cultivars identified. The study reported a moderate genetic diversity in Indian soybean cultivars and findings would be useful to the soybean breeders in selecting genetically distinct parents for a soybean improvement program.     

Detection,Localization and Tyrosine Phosphorylation Status of Ser/Thr Protein Phosphatase1γ in Freshly Ejaculated,In Vitro Capacitated and Cryopreserved Buffalo Spermatozoa

Several recent studies have indicated the important roles of Ser/Thr protein phosphatase1γ (PP1γ) in regulating the motility and capacitation of mammalian spermatozoa. Here, we report the presence and distribution of PP1γ protein in freshly ejaculated, in vitro capacitated and cryopreserved buffalo spermatozoa. The presence of PP1γ and its distribution were assessed by Western blotting and indirect immunofluorescence techniques, whereas the isoforms of PP1γ and their tyrosine phosphorylation status were identified by using 2D electrophoresis. The number of isoforms and the status of tyrosine phosphorylation of PP1γ were increased in capacitated spermatozoa when compared with freshly ejaculated spermatozoa. Differential pattern of expression and tyrosine phosphorylation of PP1γ were observed in cryopreserved spermatozoa, wherein some isoforms were degraded and some were tyrosine phosphorylated. In addition, immunofluorescence technique revealed that PP1γ was localized to principle, mid‐piece, post‐acrosomal and equatorial regions of buffalo spermatozoa. Differential distribution of tyrosine‐phosphorylated proteins were observed in fresh, capacitated and cryopreserved spermatozoa. The tyrosine phosphorylation of several proteins (20, 37, 38, 52, 60, 79 and 100 kDa) were increased when sperm cells were incubated with PP1γ inhibitor, okadaic acid. Together, our results suggest that buffalo spermatozoa express different isoforms of PP1γ protein. The protein expression and tyrosine phosphorylation of PP1γ were increased during capacitation. Furthermore, the differential pattern of expression and tyrosine phosphorylation of PP1γ were observed in cryopreserved spermatozoa. In addition, the inhibition of PP1γ protein increases protein tyrosine phosphorylation in capacitation.     

不同种植密度和施肥水平对大豆籽粒品质的影响

本试验自2002年开始至2004年止，从种植密度和施肥水平两方面对大豆脂肪、蛋白质含量的影响进行了分析。结果表明：种植密度对大豆籽粒的脂肪、蛋白质含量具有显著的影响，相关性分析结果显示，种植密度与大豆籽粒的脂肪含量存在着负相关性，与籽粒中的蛋白质含量之间为正相关。中国品种的脂肪含量受种植密度的影响大于美国品种，而中国品种的蛋白质含量受种植密度的影响小于美国品种。蛋脂总含量受种植密度的影响较小，并且不存在品种间的差异性。磷酸二铵的施用量与大豆籽粒脂肪含量和蛋白质含量之间没有显著的相关性，但是中美品种的品质对磷酸二铵施用量的反应不同。磷酸二铵的施用量与中国品种的脂肪含量、蛋白质含量和蛋脂总含量呈正相关，而与美国品种的脂肪、蛋白质含量和蛋脂总含量呈负相关，但相关程度均不显著。     

Fruit growth,cuticle deposition,water uptake,and fruit cracking in jostaberry,gooseberry, and black currant

Fruit cracking after rain limits the production of a number of crops, including some Ribes species. To gain a better understanding of the factors involved in cracking, fruit growth, deposition of the cuticular membrane (CM), water uptake and fruit cracking were studied in black currant (Ribes nigrum L. cv. Zema), gooseberry (Ribes uva-crispa L. cv. Rote Triumph), and jostaberry (Ribes nidigrolaria B. cv. Jostine). Fruit surface area and fresh mass increased continuously throughout development, whereas deposition of the CM was biphasic. CM mass per fruit increased rapidly up to 42, 41, and 49 days after full bloom (DAFB) in black currant, gooseberry, and jostaberry, respectively. Thereafter, CM mass per fruit remained constant in gooseberry and jostaberry or increased at a lower rate in black currant. The cessation of or reduced rate of CM deposition resulted in a decrease in CM mass per unit area in all berries. Elastic strain of the CM at maturity averaged 23.8% and 19.5% in gooseberry and jostaberry, respectively, and only 8.2% in black currant. Microcracks in the CM were observed first in gooseberry and jostaberry 64 DAFB, whereas there were no microcracks in black currant. Water uptake into mature detached berries was linear over 2 h of incubation. Rates of uptake were highest in gooseberry followed by black currant and jostaberry. Relative uptake was similar via the cut end of the pedicel (32.1%), the apex of the fruit (34.7%) and the fruit surface (33.2%). Rates of water uptake through the fruit surface were positively related to surface area. Average fruit water potential for black currant, gooseberry, and jostaberry was −2.14 ± 0.17, −1.24 ± 0.03, and −1.89 ± 0.20 MPa, while the permeability for osmotic water uptake was 7.7 ± 0.4 × 10⁻⁸, 5.2 ± 0.1 × 10⁻⁸, and 3.3 ± 0.3 × 10⁻⁸ m s⁻¹. Incubating whole fruit in deionized water for 72 h resulted in more cracked jostaberries (94%) than black currants (74%) or gooseberries (50%). A comparison of our findings in Ribes berries with published data for the sweet cherry drupe revealed that the berries fitted the relationships established in sweet cherry among fruit growth, cuticle deposition, strain of the cuticle, microcracking, permeability for osmotic water uptake, frequency of stomata and cracking. The Ribes berries were less susceptible to cracking than sweet cherry.