Cajanus scarabaeoides (L.), Thouars, is the closest wild crop relative of cultivated pigeonpea, Cajanus cajan (L.), Millspaugh. This wild pigeonpea has several insect-resistance mechanisms, particularly to Helicoverpa armigera (Hübner). Estimated economic losses in the semi-arid tropics from H. armigera damage in pigeonpea are approximately two billion USD/year. Therefore, it is imperative to improve pest resistance in this crop. In this study, we investigated insect-resistance components in IBS 3471, a C. scarabaeoides accession, and explored the possibility of transferring resistance mechanism/s to cultivated pigeonpea. A detached leaf bioassay revealed that IBS 3471 has more effective antibiosis and antixenosis resistance mechanisms against H. armigera compared to the susceptible C. cajan variety, ICPL 87. To further investigate the antibiosis resistance mechanism, we fed H. armigera larvae a heated and non-heated artificial diet supplemented with lyophilised IBS 3471 leaf powder. Incorporation of IBS 3471 leaf powder inhibited H. armigera larval weight and delayed larval development compared to larvae reared on diet supplemented with ICPL 87 leaf powder. The putative insect-resistance compounds in C. scarabaeoides were heat-labile. Proteomic analysis revealed higher levels of potential insecticidal proteins, namely lectin and cysteine proteinase inhibitor, in wild pigeonpea compared to the cultivated variety. Nutritional analysis and interspecific hybridisation experiments also indicated that IBS 3471 is a potential candidate for improvement of insect-resistance in pigeonpea. This study demonstrates that IBS 3471 has multiple resistance mechanisms against H. armigera, and they are transferable to cultivated pigeonpea.

Insect pests pose a serious threat to global food production. Pod borer (Helicoverpa armigera (Hübner)) is one of the most destructive pests of leguminous crops. The use of host resistance has been an effective, environmentally friendly and sustainable approach for controlling several agricultural pests. The exploitation of natural variations in crop wild relatives could yield pest-resistant crop varieties. In this study, we used a high-throughput transcriptome profiling approach to investigate the defense mechanisms of susceptible cultivated and tolerant wild pigeonpea genotypes against H. armigera infestation. The wild genotype displayed elevated pest-induced gene expression, including the enhanced induction of phytohormone and calcium/calmodulin signaling, transcription factors, plant volatiles and secondary metabolite genes compared to the cultivated control. The biosynthetic and regulatory processes associated with flavonoids, terpenes and glucosinolate secondary metabolites showed higher accumulations in the wild genotype, suggesting the existence of distinct tolerance mechanisms. This study provides insights into the molecular mechanisms underlying insect resistance in the wild pigeonpea genotype. This information highlights the indispensable role of crop wild relatives as a source of crucial genetic resources that could be important in devising strategies for crop improvement with enhanced pest resistance.

Summary: Bananas (Musa spp.) are a major fruit crop worldwide, with the Cavendish cultivar dominating the export industries, which are based primarily in the Americas and the Philippines. The sustainability of banana production in these, and other regions, is under threat from the fungal disease Fusarium wilt tropical race 4 (TR4) which kills Cavendish and many other banana cultivars. No effective TR4 control strategies exist, and no consumer‐acceptable TR4‐resistant Cavendish replacements are currently available. Previously, we identified four genetically modified (GM) TR4‐resistant Cavendish (cv. Grand Nain) banana lines (RGA2‐2, ‐3, ‐4, and ‐5), each transformed with the MamRGA2 resistance gene derived from a TR4‐resistant wild banana. Here, we provide a comprehensive agronomic evaluation of these lines and report the regulatory approval of the top‐performing line, RGA2‐4 (QCAV‐4), in Australia. After five crop cycles of field trialling, TR4 disease incidence reached 66% and 84% in the Grand Nain and Williams non‐GM controls, respectively, while GM lines showed a significantly reduced incidence ranging from 2% (QCAV‐4) to 36% (RGA2‐5). Agronomic comparisons between QCAV‐4 and non‐GM Grand Nain controls for bunch weight, yield and cycle time indicated that, for most crop cycles, there were no differences, showing that, apart from TR4 resistance, QCAV‐4 is agronomically and phenotypically comparable to conventional Grand Nain. Molecular characterization of QCAV‐4 confirmed a single, large T‐DNA insert containing multiple copies of the MamRGA2 gene on chromosome 6 of the QCAV‐4 genome. QCAV‐4 is the first GM banana approved for commercial cultivation and consumption.

Pigeonpea [Cajanus cajan (L.) Millspaugh] is an economically important legume playing a crucial role in the semi-arid tropics. Pigeonpea is susceptible to Helicoverpa armigera (Hübner), which causes devastating yield losses. This pest is developing resistance to many commercially available insecticides. Therefore, crop wild relatives of pigeonpea, are being considered as potential sources of genes to expand the genetic base of cultivated pigeonpea to improve traits such as host plant resistance to pests and pathogens. Quantitative proteomic analysis was conducted using the tandem mass tag platform to identify differentially abundant proteins between IBS 3471 and ICPL 87 tolerant accession and susceptible variety to H. armigera, respectively. Leaf proteome were analysed at the vegetative and flowering/podding growth stages. H. armigera tolerance in IBS 3471 appeared to be related to enhanced defence responses, such as changes in secondary metabolite precursors, antioxidants, and the phenylpropanoid pathway. The development of larvae fed on an artificial diet with IBS 3471 lyophilised leaves showed similar inhibition with those fed on an artificial diet with quercetin concentrations with 32 mg/25 g of artificial diet. DAB staining (3,3′-diaminobenzidine) revealed a rapid accumulation of reactive oxygen species in IBS 3471. We conclude that IBS 3471 is an ideal candidate for improving the genetic base of cultivated pigeonpea, including traits for host plant resistance.

Cajanus scarabaeoides (L.) Thouars is the closest wild relative of cultivated pigeonpea, Cajanus cajan (L.) Millspaugh. However, unlike cultivated pigeonpea which is very susceptible to insects, especially Helicoverpa armigera (Hubner), this wild pigeonpea is strongly insect resistant. Since H. armigera causes damage to many important crops resulting in economic losses up to 2 billion USD/year, improvement in resistance to this insect in crops is highly desirable. Here we investigate insect resistance components in C. scarabaeoides and explore the possibility of transferring one or more of those factors to cultivated pigeonpea. A detached leaf assay was used to assess antibiosis and antixenosis resistance mechanisms in C. scarabaeoides. Artificial diet supplemented with lyophilised leaf powder was employed to investigate the antibiosis resistance mechanism. Data on larval mortality and larval and pupal weights were collected. Time taken for neonate larvae to pupate and for pupae to develop to the moth stage were determined through daily observations. Preliminary results showed that H. armigera larval weight was significantly reduced and larval development stages were prolonged when reared on different accessions of C. scarabaeoides as compared to the susceptible check, C. cajan (ICPL 87). Interspecific hybridization was carried out between C. scarabaeoides and a commercial pigeonpea variety. Results from insect challenge assays and trichome identification indicate that the hybrids have similar levels of insect resistance to their wild parent. Proteomic analysis is being used to identify possible antibiosis factors.

Dawit, Abigail Ngugi (2021) Improvement of Helicoverpa armigera resistance in pigeonpea (Cajanus cajan) through 'omics and breeding. PhD by Publication, Queensland University of Technology.