Towards an objective evaluation of persistency of <Emphasis Type="Italic">Lolium perenne</Emphasis> swards using UAV imagery

Perennial ryegrass (Lolium perenne) is a perennial crop used in temperate regions as forage. In L. perenne breeding programs, persistency is an important trait. Poor persistency results in sward degradation and associated yield and nutritive value losses. Breeders assess persistency of accessions using visual scoring in field plots during the 2nd or 3rd growing season. This evaluation system is easy and cheap but is not free from human bias. In this study, the correlation between the scoring done by different breeders was only 0.243. As an alternative we have developed a methodology to assess persistency of L. perenne breeding materials based on vegetation indices (VIs) derived from Unmanned Aerial Vehicle (UAV) imagery. The VIs Excess green (ExG2), Green Leaf Index and Normalized green intensity (GCC) were found to provide consistent results for flights carried out under different light conditions and were validated by ground reference information. The correlation between the VIs and the percentage of ground cover extracted from on-ground imagery was 0.885. To test the implementation of the method we compared the ExG2 value based approach to selection with a visual score based selection methodology as applied by two breeders. The breeding decisions of Breeder A agreed well with decisions based on ExG2 values (74.6%), but those of Breeder B displayed a lower agreement (54.0%). In contrast, agreement between decisions based on different flights was very high (91.6%). The methodology was validated for general applicability. In summary, the results demonstrate that basing persistency selection in L. perenne breeding programs on ExG2 values from UAV imagery is likely to be more objective in comparison to the currently-used visual scoring method.     

Genetic variability of faba bean genotypes for chocolate spot (<Emphasis Type="Italic">Botrytis fabae</Emphasis>) resistance and yield

Faba bean (Vicia faba L.) has high utility as a food and soil fertility improving crop. One of the major fungal pathogens of faba bean is Botrytis fabae, the causative agent of chocolate spot. The disease affects significantly the leaf, stem, pod and seed of faba bean compromise its productivity in the smallholder farming sector. Nonetheless, there are limited resistant/tolerant faba bean varieties available and disease control technology options. Therefore, it was prudent to evaluate faba bean landraces for chocolate spot resistance. Fifty landraces together with ten improved varieties were evaluated both in the field and in the greenhouse under natural and artificial inoculation with previously selected aggressive Botrytis fabae isolate (Iso-016) from West Gojjam, in Ethiopia. There were highly significant differences (p?<?0.001) among the landraces for reaction to the disease and agronomic traits. Significant positive correlation was recorded between reaction of genotypes in the field and greenhouse disease data. The overall mean disease epidemics varied from 92.5 to 697.5 for the area under disease progress curve (AUDPC). The highest level of resistance was found in the ICARDA lines, ILB-4726, ILB-938 and BPL-710. Of all 18 landrace collections displayed significantly lower disease reaction than the susceptible check. However the resistance was moderate. The selected eighteen landraces will be recommended for use in breeding for chocolate resistance. Overall, resistance was highly heritable, suggesting that phenotypic selection can be exploited to improve chocolate spot resistance in faba bean varieties.     

Identification of drought tolerant populations at multi-stage growth phases in temperate maize germplasm

Drought is one of the main abiotic stresses in agriculture worldwide. Drought could increase under the predicted scenario of climate change, particularly in the Mediterranean area. Breeding for drought tolerance requires screening of germplasm in order to identify sources of tolerance; therefore, were evaluated 51 diverse open-pollinated maize populations from various temperate regions under increasing levels of drought at germination, seedling establishment and early growth. There was genetic variability for drought tolerance among populations at all growth phases. Several populations from diverse origins and germplasm groups exhibited high germination across stress treatments. Some of those populations had high ability to sustain root development and showed differential performance depending on the stage of development and the phenotypic aspect considered. In general, BS17 showed high germination rate, fast seedling growth and early vigor under drought. Longlellow and Grano de trigo showed high germination and growth establishment rates, whereas AS3(HT)C3 showed high germination rate and early vigor under drought. The photosynthetic rate, stomatal conductance and transpiration of Enano Levantino/Hembrilla (ELH) and BS17 populations were not affected by drought. Water use efficiency of BS17, ELH, Northwestern Dent (NWD) and Viana was not affected by drought. Some of these populations are promising sources of drought tolerance which can provide different mechanisms of drought tolerance at different stages of plant development. Therefore, these results open new possibilities of breeding for drought tolerance by combining those mechanisms through crosses among potential donors. Furthermore, these findings indicate that it is worthwhile to study the genetic and biological basis of such mechanisms.     

Are tree breeders properly predicting genetic gain? A case study involving <Emphasis Type="Italic">Corymbia</Emphasis> species

The estimation of quantitative genetic parameters in breeding programs is important to ensure efficient selection. In this context, knowledge of the mating system is critical, as it underpins assumptions about inter-relatedness on which variance component estimation depends. However, proper account of the breeding system is not always taken, either because it is unknown and/or because it is ignored. That eucalypts have a mixed-mating system is well-established, however many breeders use models that assume outcrossed mating with an infinite number of male parents (i.e. allogamous mating), from which genetic parameter estimates are then used to predict genetic gains. First-generation, open-pollinated progeny tests of Corymbia citriodora subsp. citriodora and C. citriodora subsp. variegata, being managed for seed production, were used to investigate the likely bias, resulting in overestimation of genetic parameters under the assumption of allogamous mating. When we assumed allogamous mating, we observed inflated predictions of additive variance and narrow-sense heritability (\(\hat{h}_{a}^{2}\)) of diameter at breast height and height. The overestimate of \(\hat{h}_{a}^{2}\) was approximately 32% for C. citriodora subsp. citriodora and 21% for C. citriodora subsp. variegata. Inappropriate modelling of relatedness in eucalypts that assumes panmictic outcrossing when in fact these species have a mixed-mating system results in overestimates of the population genetic gain with selection.     

Evaluation of a multi-parent advanced generation inter-cross (MAGIC) introgressed line population for Verticillium wilt resistance in Upland cotton

Verticillium wilt (VW, caused by Verticillium dahliae Kleb) is a destructive fungal soil-borne disease in Upland cotton (Gossypium hirsutum L.). High levels of VW resistance can be transferred into Upland from Pima cotton (G. barbadense L.) through interspecific introgression breeding. In this greenhouse study, VW resistance was evaluated in a multi-parent advanced generation inter-cross (MAGIC) introgressed line (IL) population, derived from a random mated Barbadense Upland population with five generations of intermating (called RMBUP-C4) between three Upland cotton cultivars and 18 CS-B Upland lines each carrying a pair of G. barbadense chromosome or arm in the TM-1 background. The objectives of this study were to, (1) evaluate VW resistance of 530 MAGIC ILs in the greenhouse; and (2) to identify lines with VW resistance in the MAGIC population based on a total of three replicated greenhouse tests. Approximately 8 plants for each line in each replicate were grown and screened for VW resistance using three parameters i.e., disease leaf severity rating, percentage defoliated leaves, and percentage infected plants, with a total of ~?25,190 plants evaluated. A correlation analysis indicated that the three parameters were significantly and positively correlated with one another in each test. The disease leaf severity rating was the best parameter to assess VW resistance due to its relatively low coefficient of variation and its higher resolution to differentiate resistant genotypes from susceptible ones. Of the 530 genotypes, 5 showed resistance to VW, namely, NMIL348, NMIL518, NMIL405, NMIL290, NMIL307 and had higher levels of resistance to VW with mean disease leaf severity ratings, percentage of defoliated leaves, and percentage of infected plants across three tests ranging from 0.58–1.46, 9.46–26.74, and 25–95%, respectively. These lines can be used as parental lines to improve VW resistance in cotton breeding programs.     

Pollen characteristics and stigma receptivity for <Emphasis Type="Italic">Anemone coronaria</Emphasis> L.

To obtain crossing products, an efficient pollination and subsequent fertilization is essential. This efficient pollination is achieved by pollen germination and tube growth. Here, these pollen characteristics of 2 genetically differentiating cultivars of Anemone coronaria L. were investigated in vitro. For the essential components boron, calcium and an osmoticant, only calcium showed to be crucial for pollen germination. Boron concentrations influenced the pollen tube length with a concentration of 100 mg l^?1 H₃BO₃ resulting in the longest pollen tubes. For the osmoticant sucrose, a concentration of 100 g l^?1 caused a significant positive effect on both pollen germination and pollen tube length for the 2 cultivars. The cultivars reacted similarly with respect to the investigated compounds. Next to this, the pollen development was correlated with 8 different flower stages in A. coronaria. As a final point, the germination of an optimized pollen germination medium was compared with in vivo pollen germination in cross-pollinations within the same cultivar (identified by aniline blue staining). For ‘Mistral Wine’, pollen germination percentage was lower in vitro than in vivo, while ‘Wicabri Blue’ pollen showed no significant difference in germination rates in vivo and in vitro. To achieve fertilization a following requisite is that the stigma is receptive. To study this, the most receptive female flower stage of the 8 different flower stages for A. coronaria was characterized by aniline blue staining. A. coronaria clearly showed protogyny.     

Identification of PCR-based DNA Marker Linked to High Phytase Level of Wheat

The phytase is a key enzyme to hydrolyze phytic acid present in wheat grains and improves the bio-availability of micronutrients in monogastric animals. Phytase trait being contributed by specific regions of the genome requires identification of these regions, using suitable molecular markers. Hence, in the present investigation we attempted to develop a PCR-based marker that detects the phytase level in wheat. Six sets of PCR primers were designed on the basis of nucleotides sequence variation found in the sequence of both varieties. Out of six set of primers, one set amplified two different sized bands, i.e. 334 bp and 295 in two wheat cultivars C-306 (low phytase) and DBW 17 (high phytase), respectively. It exhibited a polymorphic banding pattern with length polymorphism and clearly separating low and high phytase genotypes. The primer set was also used for PCR of 46 synthetic hexaploids and 46 release varieties of wheat to validate the developed markers. Association among identified markers and phytase activity was found to be at 99.9% confidence level based on Fisher’s exact test (F-test). Therefore, this PCR primer set will be useful to select the wheat germplasm having high phytase levels and also in wheat breeding programs aimed at improving phytase levels in bread wheat cultivars.     

Changes in the Functional Components and Radical Scavenging Activity of Korean Maize Hybrids According to Different Cropping Seasons

We evaluated the proximate composition, free sugar content, fatty acid composition, carotenoid content, total phenol content, and radical scavenging activity of the grain from various Korean maize hybrid cultivars grown in two different cropping seasons. The moisture, crude ash, crude fat, crude protein, total starch, and amylose contents were significantly higher in most of the maize hybrid cultivars when grown in the early-season than when grown in the late-season. The free sugar content, fatty acid composition, carotenoid content, and total phenol content differed significantly between cultivars and cropping seasons. The highest unsaturated fatty acid compositions of maize hybrids of early-season and late-season were 86.05 and 86.29%, respectively, in the Daanok cultivar. The carotenoid contents were significantly higher in maize hybrids of late-season compared to those of early-season. The highest total phenol content was 108.09 mg/100 g in Singwangok of the late-season. The radical scavenging activity of maize hybrids differed significantly between cultivars and cropping seasons.     

Evaluation of Seed Vigor Tests for Predicting Seedling Establishment at Low Temperature in Rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

This study was carried out to develop an evaluation method to predict rice seedling establishment (SE) under low-temperature conditions. Two Korean-bred japonica cultivars, Shindongjin and Hopum were used in the experiment. Fresh seeds were treated with an accelerated aging (AA) at 40°C and 100% RH for 1-15 days. The SEs of the fresh and AA seeds were evaluated in nursery beds at 17°C, and their correlation coefficients with seed vigor values measured by 9 test methods including standard germination test (SGT), cool germination test (CLT), cold germination test (CDT), seedling growth rate test (SGRT), ascorbate peroxidase (APX), peroxidase (POX), superoxide dismutase (SOD), catalase (CAT), and α ‐amylase (AMY) activities. The percentage of SE decreased slowly from 75 to 0% with an increasing of AA period from 0 to 15 days. The result of nine vigor tests showed different correlations with the SE. SGT, CLT, SGRT, and POX were significantly correlated with the SE. In the correlation analysis with only short-term aging seeds (1-7 days), the SE was very highly correlated with SGT, CLT, CDT, SGRT, POX, and CAT. These results suggest that seed vigor values measured by several methods including SGT and POX could be used as a reference value to secure SE at low temperatures in nursery bed rice seedling culture.