CRISPR/Cas9-gene editing approaches in plant breeding.

CRISPR/Cas9 gene editing system is recently developed robust genome editing technology for accelerating plant breeding. Various modifications of this editing system have been established for adaptability in plant varieties as well as for its improved efficiency and portability. This review provides an in-depth look at the various strategies for synthesizing gRNAs for efficient delivery in plant cells, including chemical synthesis and in vitro transcription. It also covers traditional analytical tools and emerging developments in detection methods to analyze CRISPR/Cas9 mediated mutation in plant breeding. Additionally, the review outlines the various analytical tools which are used to detect and analyze CRISPR/Cas9 mediated mutations, such as next-generation sequencing, restriction enzyme analysis, and southern blotting. Finally, the review discusses emerging detection methods, including digital PCR and qPCR. Hence, CRISPR/Cas9 has great potential for transforming agriculture and opening avenues for new advancements in the system for gene editing in plants.

Effect of mother bonded rearing on growth, health and physiological state of Murrah buffalo calves.

We investigated the effect of different types of mother-young contacts (fenceline, restricted and no contact) on the health, growth performance, behaviour and physiological state of Murrah buffalo calves. A total of 24 calves were allocated to three groups of 8: NCM (no calf-mother contact), RCM (restricted calf-mother contact) and FCM (fenceline calf-mother contact). At three months of age, the FCM calves had a higher average body weight (58.6 ± 1.5 kg) than the RCM (52.8 ± 1.3 kg) and NCM (53.6 ± 1.3 kg) calves (P < 0.05). The average daily gain (ADG) at three months of age was greater in FCM (0.6 ± 0.1 kg/d) than RCM (0.5 ± 0.1 kg/d) and NCM (0.5 ± 0.1 kg/d) calves (P < 0.05). The mean immunoglobulin G (IgG) concentrations were significantly greater on d 7, 28, 42 and d 56 of sampling (P < 0.05 for all comparisons) in FCM than RCM and NCM calves. The mean cortisol levels were highest in the NCM calves followed by RCM and the lowest values were observed in FCM calves on d 0, d 7, d 28, d 42 and d 56, respectively (P < 0.05, for all recorded days). The diarrhoea score and eggs per gram of faeces (EPG) were significantly greater in the NCM group compared to the RCM and FCM groups. The time spent in cross sucking, licking inanimate objects and self-licking was greatest (P < 0.05) in NCM calves, followed by RCM calves, and was almost non-existent in the FCM group on all the recorded weeks. Time spent in backward ear position was greater (P < 0.05) in FCM than RCM and NCM calves. The average time spent in forward ear position was greatest (P < 0.05) in the RCM calves followed by NCM with the lowest time observed in FCM calves. The mean duration of time spent by calves in the asymmetrical and axial ear position differed significantly (P < 0.05) among NCM, RCM and FCM calves, P < 0.05. The full and fenceline buffalo calf-mother contact system showed higher growth rates, lower levels of stress responses, oral stereotypies and eggs per gram of faeces. In addition, ear postures may be used as a dependable, and reliable measure of positive, low arousal emotional states.