Discrimination of haploid and diploid maize kernels via multispectral imaging

The use of doubled haploids (DHs) in maize has become ubiquitous in maize breeding programmes as it allows breeders to go from cross to evaluation in as little as 2 years. Two important aspects of the in vivo DH system used in maize are as follows: (i) the identification of haploid progeny and (ii) doubling of the haploid genome to produce fertile inbred lines. This study is focused on the first step. Currently, identification of maize haploid progeny is performed manually using the R1-nj seed colour marker. This is a labour-intensive and time-consuming process; a method for automated sorting of haploids would increase the efficiency of DH line development. In this study, six inbred lines were crossed with the maternal haploid inducer ‘RWS/RWK-76’ and a sample of seed was sorted manually for each line. Using the VideometerLab 3 system, spectral imaging techniques were applied to discriminate between haploids and hybrids. Using DNA markers to confirm the haploid/diploid state of the tested seed, for the majority of genotypes haploid identification was possible with over 50% accuracy.

[1]  René Gislum,et al.  Use of partial least squares discriminant analysis on visible‐near infrared multispectral image data to examine germination ability and germ length in spinach seeds , 2012 .

[2]  B. L. Welch.  THE SIGNIFICANCE OF THE DIFFERENCE BETWEEN TWO MEANS WHEN THE POPULATION VARIANCES ARE UNEQUAL , 1938 .

[3]  Thomas Lübberstedt,et al.  Weighing in on a method to discriminate maize haploid from hybrid seed , 2015 .

[4]  S. S. Chase.  Monoploid Frequencies in a Commercial Double Cross Hybrid Maize, and in Its Component Single Cross Hybrids and Inbred Lines. , 1949, Genetics.

[5]  Hartwig H. Geiger,et al.  In vivo haploid induction in maize - performance of new inducers and significance of doubled haploid lines in hybrid breeding , 2005 .

[6]  Albrecht E. Melchinger,et al.  In Vivo Haploid Induction in Maize: Identification of Haploid Seeds by Their Oil Content , 2014 .

[7]  Roger W. Jones,et al.  Selection of Haploid Maize Kernels from Hybrid Kernels for Plant Breeding Using Near-Infrared Spectroscopy and SIMCA Analysis , 2012, Applied spectroscopy.

[8]  A. Melchinger,et al.  Oil Content is Superior to Oil Mass for Identification of Haploid Seeds in Maize Produced with High‐Oil Inducers , 2015 .

[9]  E. Coe.  A Line of Maize with High Haploid Frequency , 1959, The American Naturalist.

[10]  Prasanna M. Boddupalli,et al.  Analysis of effectiveness of R1-nj anthocyanin marker for in vivo haploid identification in maize and molecular markers for predicting the inhibition of R1-nj expression , 2014, Theoretical and Applied Genetics.

[11]  J. Carstensen,et al.  Multispectral imaging as a potential tool for seed health testing of spinach (Spinacia oleracea L.). , 2011 .

[12]  Noboru Higashi,et al.  Far-Ultraviolet Spectroscopy in the Solid and Liquid States: A Review , 2012 .

[13]  C. N. Stewart,et al.  A rapid CTAB DNA isolation technique useful for RAPD fingerprinting and other PCR applications. , 1993, BioTechniques.

[14]  Changhong Liu,et al.  Non-destructive discrimination of conventional and glyphosate-resistant soybean seeds and their hybrid descendants using multispectral imaging and chemometric methods , 2014, The Journal of Agricultural Science.

[15]  Gerald N. De La Fuente,et al.  Accelerating plant breeding. , 2013, Trends in plant science.

[16]  Albrecht E. Melchinger,et al.  Doubled Haploids in Tropical Maize: I. Effects of Inducers and Source Germplasm on in vivo Haploid Induction Rates , 2011 .

[17]  Basil J. Nikolau,et al.  Haploid differentiation in maize kernels based on fluorescence imaging , 2016 .

[18]  Albrecht E. Melchinger,et al.  Rapid and accurate identification of in vivo-induced haploid seeds based on oil content in maize , 2013, Scientific Reports.