Emerging technological developments to address pest resistance in Bt cotton

Cotton plays a crucial role in shaping Indian economy and rural livelihoods.The cotton crop is prone to numerous insect pests,necessitating insecticidal application,which increases production costs.The advent of the expression of Bacillus thuringiensis(Bt)insecticidal protein in cotton has significantly reduced the burden of pest without compromising environmental or human health.After the introduction of transgenic cotton,the cultivated area expanded to 22 million hectares,with a 64% increase in adoption by farmers worldwide.Currently,Bt cotton accounts for 93% of the cultivated cotton area in India.However,extensive use of Bt cotton has accelerated resistance development in pests like the pink bollworm.Furthermore,the overreliance on Bt cotton has reduced the use of broad-spectrum pesticides,favouring the emergence of secondary pests with significant challenges.This emphasizes the urgent necessity for developing novel pest management strategies.The high-dose and refuge strategy was initially effective for managing pest resistance in Bt cotton,but its implementation in India faced challenges due to misunderstandings about the use of non-Bt refuge crops.Although gene pyramiding was introduced as a solution,combining mono toxin also led to instances of cross-resistance.Therefore,there is a need for further exploration of biotechnological approaches to manage insect resistance in Bt cotton.Advanced biotechnological strategies,such as sterile insect release,RNA interference(RNAi)-mediated gene silencing,stacking Bt with RNAi,and genome editing using clustered regularly interspaced short palindromic repeats/CRISPR-associated protein(CRISPR-Cas),offer promising tools for identifying and managing resistance genes in insects.Additionally,CRISPR-mediated gene drives and the development of novel biopesticides present potential avenues for effective pest management in cotton cultivation.These innovative approaches could significantly enhance the sustainability and efficacy of pest resistance management in Bt cotton.

STUDY ON THE ANTI-TUMOR EFFICACY INDUCED BY HEAT SHOCK PROTEIN 70-PEPTIDE COMPLEXES DERIVED FROM TUMOR CELLS

OBJECTIVE: To study the efficacy and explore the mechanism of the anti-tumor immunity elicited by heat shock protein 70-peptide complexes (HSP70-PC) derived from tumor cells. METHODS: Cells culture, flow cytometric analysis, affinity chromatography for protein purification, SDS-PAGE, Western-blotting and animal experiment were used. RESULTS: HSP70-PC immunization rendered protective effect to both naive tumorl-bearing mice. All of the naive mice obtained complete resistance to Hcaf cell attack; 40% of the tumor-bearing mice survived for over 90 days, whereas the mice of control group died within 2 weeks (P

Effect of dihydrofolate reductase gene knock-down on the expression of heart and neural crest derivatives expressed transcript 2 in zebrafish cardiac development

Background Folic acid is very important for embryonic development and dihydrofolate reductase is one of the key enzymes in the process of fotic acid performing its biological function. Therefore, the dysfunction of dihydrofolate reductase can inhibit the function of folic acid and finally cause the developmental malformations. In this study, we observed the abnormal cardiac phenotypes in dihydrofolate reductase （DHFR） gene knock-down zebrafish embryos, investigated the effect of DHFR on the expression of heart and neural crest derivatives expressed transcript 2 （HAND2） and explored the possible mechanism of DHFR knock-down inducing zebrafish cardiac malformations. Methods Morpholino oligonucleotides were microinjected into fertilized eggs to knock down the functions of DHFR or HAND2. Full length of HAND2 mRNA which was transcribed in vitro was microinjected into fertilized eggs to overexpress HAND2. The cardiac morphologies, the heart rates and the ventricular shortening fraction were observed and recorded under the microscope at 48 hours post fertilization. Whole-mount in situ hybridization and real-time PCR were performed to detect HAND2 expression. Results DHFR or HAND2 knock-down caused the cardiac malformation in zebrafish. The expression of HAND2 was obviously reduced in DHFR knock-down embryos （P〈0.05）. Microinjecting HAND2 mRNA into fertilized eggs can induce HAND2 overexpression. HAND2 overexpression rescued the cardiac malformation phenotypes of DHFR knock-down embryos. Conclusions DHFR plays a crucial role in cardiac development. The down-regulation of HAND2 caused by DHFR knock-down is the possible mechanism of DHFR knock-down inducing the cardiac malformation.