Application of commercially available, low-cost, miniaturised NIR spectrometers to the assessment of the sugar content of intact fruit

Recent decreases in costs, and improvements in performance, of silicon array detectors open a range of potential applications of relevance to plant physiologists, associated with spectral analysis in the visible and short-wave near infra-red (far-red) spectrum. The performance characteristics of three commercially available ‘miniature’ spectrometers based on silicon array detectors operating in the 650–1050-nm spectral region (MMS1 from Zeiss, S2000 from Ocean Optics, and FICS from Oriel, operated with a Larry detector) were compared with respect to the application of non-invasive prediction of sugar content of fruit using near infra-red spectroscopy (NIRS). The FICS–Larry gave the best wavelength resolution; however, the narrow slit and small pixel size of the charge-coupled device detector resulted in a very low sensitivity, and this instrumentation was not considered further. Wavelength resolution was poor with the MMS1 relative to the S2000 (e.g. full width at half maximum of the 912 nm Hg peak, 13 and 2 nm for the MMS1 and S2000, respectively), but the large pixel height of the array used in the MMS1 gave it sensitivity comparable to the S2000. The signal-to-signal standard error ratio of spectra was greater by an order of magnitude with the MMS1, relative to the S2000, at both near saturation and low light levels. Calibrations were developed using reflectance spectra of filter paper soaked in range of concentrations (0–20% w/v) of sucrose, using a modified partial least squares procedure. Calibrations developed with the MMS1 were superior to those developed using the S2000 (e.g. coefficient of correlation of 0.90 and 0.62, and standard error of cross-validation of 1.9 and 5.4%, respectively), indicating the importance of high signal to noise ratio over wavelength resolution to calibration accuracy. The design of a bench top assembly using the MMS1 for the non-invasive assessment of mesocarp sugar content of (intact) melon fruit is reported in terms of light source and angle between detector and light source, and optimisation of math treatment (derivative condition and smoothing function).

Seasonal effects of foliar application of phosphonate on phosphonate translocation, in vitro pollen viability and pollen germination in `Hass' avocado (Persea americana Mill.)

Phosphonate fungicides are used widely in the control of diseases caused by Phytophthora cinnamomi Rands. For the most part phosphonate is seen as a safe to use on crops with phytotoxicity rare. However, recent research has shown that phosphonate has detrimental effects on the floral biology of some indigenous Australian plants. Since phosphonate fungicides are regularly used for the control of Phytophthora root rot in avocados, research was carried out to study the translocation of phosphonate fungicide in 'Hass' trees and any effects on their floral biology. Field-grown trees were sprayed with 0, 0.06 or 0.12 M mono-dipotassium phosphonate (pH 7.2) at summer flush maturity, floral bud break or anthesis. Following treatment, phosphonic acid concentrations were determined in leaves, roots, inflorescence rachi and flowers and in vitro pollen germination and pollen tube growth studied. Phosphonic acid concentration in the roots and floral parts was related to their sink strength at the respective times of application with concentration in roots highest (36.9.mg g±1) after treatment at summer flush maturity and in flowers (234.7 mg g±1) after treatment during early anthesis. Phosphonate at >0.03 M was found to be significantly phytotoxic to in vitro pollen germination and pollen tube growth. However, this rate gave a concentration far in excess of that measured in plant tissues following standard commercial applications of mono-dipotassium phosphonate fungicide. There was a small effect on pollen germination and pollen tube growth when 0.06 and 0.12 M mono-dipotassium phosphonate was applied during early anthesis. However, under favourable pollination and fruit set conditions it is not expected to have commercial impact on tree yield. However, there may be detrimental commercial implications from phosphonate sprays at early anthesis if unfavourable climatic conditions for pollination and fruit set subsequently occur. A commercial implication from this study is that phosphonic acid root concentrations can be elevated and maintained with strategic foliar applications of phosphonate fungicide timed to coincide with peaks in root sink strength. These occur at the end of the spring and summer flushes when shoot growth is relatively quiescent. Additional foliar applications may be advantageous in under high disease-pressure situations but where possible should be timed to minimize overlap with other significant growth events in the tree such as rapid inflorescence, and fruit development and major vegetative flushing.

Development of near infrared analysis of faeces to estimate non-grass proportions in diets selected by cattle grazing tropical pastures

Grass (monocots) and non-grass (dicots) proportions in ruminant diets are important nutritionally because the non-grasses are usually higher in nutritive value, particularly protein, than the grasses, especially in tropical pastures. For ruminants grazing tropical pastures where the grasses are C-4 species and most non-grasses are C-3 species, the ratio of C-13/C-12 in diet and faeces, measured as delta C-13 parts per thousand, is proportional to dietary non-grass%. This paper describes the development of a faecal near infrared (NIR) spectroscopy calibration equation for predicting faecal delta C-13 from which dietary grass and non-grass proportions can be calculated. Calibration development used cattle faeces derived from diets containing only C-3 non-grass and C-4 grass components, and a series of expansion and validation steps was employed to develop robustness and predictive reliability. The final calibration equation contained 1637 samples and faecal delta C-13 range (parts per thousand) of [12.27]-[27.65]. Calibration statistics were: standard error of calibration (SEC) of 0.78, standard error of cross-validation (SECV) of 0.80, standard deviation (SD) of reference values of 3.11 and R-2 of 0.94. Validation statistics for the final calibration equation applied to 60 samples were: standard error of prediction (SEP) of 0.87, bias of -0.15, R-2 of 0.92 and RPD of 3.16. The calibration equation was also tested on faeces from diets containing C-4 non-grass species or temperate C-3 grass species. Faecal delta C-13 predictions indicated that the spectral basis of the calibration was not related to C-13/C-12 ratios per se but to consistent differences between grasses and non-grasses in chemical composition and that the differences were modified by photosynthetic pathway. Thus, although the calibration equation could not be used to make valid faecal delta C-13 predictions when the diet contained either C-3 grass or C-4 non-grass, it could be used to make useful estimates of dietary non-grass proportions. It could also be ut :sed to make useful estimates of non-grass in mixed C-3 grass/non-grass diets by applying a modified formula to calculate non-grass from predicted faecal delta C-13. The development of a robust faecal-NIR calibration equation for estimating non-grass proportions in the diets of grazing cattle demonstrated a novel and useful application of NIR spectroscopy in agriculture.

New SNPs for population genetic analysis reveal possible cryptic speciation of eastern Australian sea mullet (Mugil cephalus)

Sustainable management of sea mullet (Mugil cephalus) fisheries needs to account for recent observations of regional-scale differentiation. Population genetic analysis is sought to assess the situation of this ecologically and economically important fish species in eastern Australian waters. Here, we report (i) new population genetic markers [single nucleotide polymorphisms (SNPs) and potential microsatellites], (ii) first estimates of spatial genetic differentiation and (iii) prospective power tests for designing more comprehensive studies. Six DNA samples from three sampling regions (North Queensland, South Queensland and central New South Wales) on the eastern coast of Australia were used to prepare restriction site associated DNA (RAD) tag libraries from genomic DNA digested with EcoRI and MseI. A pooled sample of regional RAD tag libraries was sequenced using the Roche GS-FLX Titanium platform. A total of 172837 raw reads (17.4Mbp) were retrieved, 95500 of which were used to discover 1267 SNPs and 1417 microsatellites. A subset of 161 SNPs was validated based on 63 additional DNA samples genotyped using the Sequenom MassArray (iPLEX Gold chemistry). Altogether 92 SNPs (57%) were confirmed, with 40% of these marking fixed variants between northern and southern sampling regions. Our preliminary findings indicate a multispecies fishery stock of M. cephalus in eastern Australian waters, but suggest that strong genetic differentiation occurs north of major fishing grounds. Low potential differentiation within major fishing grounds (e.g. FST=0.0025) can be resolved with a likely power 67% by using standard sample sizes of 50 and validated subsets of available markers.