Seasonal and Species Variation of the Hepatotoxin Indospicine in Australian Indigofera Legumes as Measured by UPLC−MS/MS

Livestock industries have maintained a keen interest in pasture legumes because of the high protein content and nutritive value. Leguminous Indigofera plant species have been considered as having high feeding values to be utilized as pasture, but the occurrence of the toxic constituent indospicine in some species has restricted this utility. Indospicine has caused both primary and secondary hepatotoxicosis and also reproductive losses, but has only previously been determined in a small number of Indigofera species. This paper validates a high throughput ultra-performance liquid chromatography−tandem mass spectrometry (UPLC−MS/MS) method to determine indospicine content of various Indigofera species found in Australian pasture. Twelve species of Indigofera together with Indigastrum parviflorum plants were collected and analysed. Out of the 84 samples analyzed, *I. spicata contained the highest indospicine level (1003 ± 328 mg/kg DM, n = 4) followed by I. linnaei (755 ± 490 mg/kg DM, n = 51). Indospicine was not detected in 9 of the remaining 11 species, and at only low levels (<10 mg/kg DM) in 2 out of 8 I. colutea specimens and in 1 out of 5 I. linifolia specimens. Indospicine concentrations were below quantitation levels for other Indigofera spp. (I. adesmiifolia, I. georgei, I. hirsuta, I. leucotricha,* I. oblongifolia, I. australis and I. trita) and Indigastrum parviflorum. One of the more significant findings to emerge from this study is that the indospicine content of I. linnaei is highly variable (159 to 2128 mg/kg DM, n = 51), and differs across both regions and seasons. Its first re-growth after spring rain has a higher (p < 0.01) indospicine content than growth following more substantial summer rain. The species collected include the predominant Indigofera in Australia pasture, and of these, only *I. spicata and I. linnaei contain high enough levels of indospicine to pose a potential toxic threat to grazing herbivores.

Variation in susceptibility among macadamia genotypes and species to Phytophthora root decay caused by Phytophthora cinnamomi

Phytophthora cinnamomi is a major pathogen of cultivated macadamia (Macadamia integrifolia, Macadamia tetraphylla and their hybrids) worldwide. The susceptibility of the two non-edible Macadamia species (Macadamia ternifolia and Macadamia jansenii) to P. cinnamomi is not well-understood. Commercial macadamia trees are established on grafted seedling (seed propagation) or own-rooted cutting (vegetative propagation) rootstocks of hybrids of the cultivated species. There is little information to support the preferential use of rootstock propagated by either seedling or own-rooted cutting methods in macadamia. In this study we assessed roots of macadamia plants of the four species and their hybrids, derived from the two methods of propagation, for their susceptibility to P. cinnamomi infection. The roots of inoculated plant from which P. cinnamomi was recovered showed blackening symptoms. The non-cultivated species, M. ternifolia and M. jansenii and their hybrids were the most susceptible germplasm compared with M. tetraphylla and M. integrifolia. Of these two species, M. tetraphylla was less susceptible than M. integrifolia. Significant differences were observed among the accessions of their hybrids. A strong association (R2 > 0.75) was recorded between symptomatic roots and disease severity. Root density reduced with increasing disease severity rating in both own-rooted cuttings (R2 = 0.65) and germinated seedlings (R2 = 0.55). P. cinnamomi severity data were not significantly (P > 0.05) different between the two methods of plant propagation. The significance of this study to macadamia breeding and selection of disease resistant rootstocks is discussed.