Codon usage bias (CUB) phenomenon varies with the species and even within the genes of the same species, where few codons are preferred more frequently than their other synonymous codons. It also categorizes the differences between species. Nucleotide compositional analysis reveals the molecular mechanisms of genes and the evolutionary relationship of a gene in dissimilar plant species. In the present study, three orthologous sequences of each FAE1 (FAE1.1, FAE1.2, and FAE1.3) and FAD2 (FAD2.1, FAD2.2, and FAD2.3) genes, from six Brassica species were accessed using the GenBank database. Further, CUB-related parameters such as nucleotide composition (AT and GC content), relative synonymous codon usage (RSCU), the effective number of codons (ENC), frequency of optimal codons (Fop), relative codon usage bias (RCBS), neutrality plot (GC12 vs. GC3), parity rule-2 [(A3/(A3 + T3) vs. (G3/(G3 + C3)], and correspondence analysis (COA) were analyzed to compare codon bias in U’s triangle Brassica species. The FAE1 genes were AT-biased and FAD2 genes were GC-biased across the studied Brassica species. RSCU values indicated that both the genes had moderate codon usage frequency for selected amino acids. The evolutionary study confirmed that codon usage preference is similar within the species grouped into the same cluster for FAE1; however, B. nigra performed differently for FAD2.2 orthologue. The high ENC value, low Fop, and RSCU value highlighted that FAE1 and FAD2 genes had a low level of gene expression and moderate preference for codon usage across the Brassicas. In addition, neutrality plot, parity rule, and correspondence analysis revealed that natural selection pressure had significantly contributed to CUB for FAE1 genes, whereas mutation and selection pressure occurred for FAD2 genes. This study would help to decode codon optimization, improve the level of expression of exogenous genes, and transgenic engineering to increase fatty acid profiling for the betterment of seed oil in Brassica species.

White rust [Albugo candida (Pers. Ex. Lev) Kuntze] is an important oomycetes disease of Indian mustard [Brassica juncea (L.) Czern & Coss] that causes a drastic reduction in seed yield and quality when the conditions are favorable. A set of 25 Indian mustard genotypes were screened against A. candida Delhi isolate (Ac-Dli) under both controlled and natural epiphytotic conditions. Out of 25, only six genotypes (Pusa Karishma, PDZ-3, Heera, BEC-144, BIO YSR, and Donskaja) were found highly resistant in both environments. To study the inheritance of resistance, four resistant genotypes (BEC-144, BIO YSR, Pusa Karishma, and Donskaja) were crossed with three susceptible genotypes (PM-24, Pusa Vijay, and MSTWR-17-15) in a definite design. The parents, F₁, F_2, and BC₁F₁ (F₁ backcrossed with susceptible parent) generations were screened under both artificially controlled and natural epiphytotic conditions and the results indicated that the resistance in BIO YSR, BEC-144, and Pusa Karishma is governed by a single dominant gene, but more than one dominant gene is responsible for resistance in Donskaja. A test of an allelism conducted on the F₂ population derived by crossing resistant (BEC-144) × resistant (BIO YSR) genotypes indicated that the gene imparting resistance to Ac-Dli isolate in the resistant parents BEC-144 and BIO YSR is the same and they are allelic to each other. Therefore, in broader terms, the information generated in the present study will be useful in Indian mustard breeding programs for the development of durable resistant cultivars.

In the era of global warming, heat stress, particularly at the seedling stage, is a major problem that affects the production and productivity of crops such as mustard that are grown in cooler climates. Nineteen mustard cultivars were exposed to contrasting temperature regimes—20 °C, 30 °C, 40 °C and a variable range of 25–40 °C—and evaluated for changes in physiological and biochemical parameters at the seedling stage to study their role in heat-stress tolerance. Exposure to heat stress showed detrimental effects on seedling growth as revealed by reduced vigor indices, survival percentages, antioxidant activity and proline content. The cultivars were grouped into tolerant, moderately tolerant and susceptible based on the survival percentage and biochemical parameters. All the conventional and three single-zero cultivars were found to be tolerant and moderately tolerant, respectively, while double-zero cultivars were reckoned to be susceptible except for two cultivars. Significant increases in proline content and catalase and peroxidase activities were found associated with thermo-tolerant cultivars. More efficient antioxidant system activity and proline accumulation were noticed in conventional along with three single-zero (PM-21, PM-22, PM-30) and two double-zero (JC-21, JC-33) cultivars that might have provided better protection to them under heat stress than the remaining one single- and nine double-zero cultivars. Tolerant cultivars also resulted in significantly higher values of most of the yield attributing traits. Heat-stress-tolerant cultivars could easily be selected based on the survival percentage, proline and antioxidants at the seedling stage and included as efficient cultivars in breeding programs.