Crop-Specific Responses to Cold Stress and Priming: Insights from Chlorophyll Fluorescence and Spectral Reflectance Analysis in Maize and Soybean.

This study aimed to investigate the impact of cold stress and priming on photosynthesis in the early development of maize and soybean, crops with diverse photosynthetic pathways. The main objectives were to determine the effect of cold stress on chlorophyll a fluorescence parameters and spectral reflectance indices, to determine the effect of cold stress priming and possible stress memory and to determine the relationship between different parameters used in determining the stress response. Fourteen maize inbred lines and twelve soybean cultivars were subjected to control, cold stress, and priming followed by cold stress in a walk-in growth chamber. Measurements were conducted using a portable fluorometer and a handheld reflectance instrument. Cold stress induced an overall downregulation of PSII-related specific energy fluxes and efficiencies, the inactivation of RCs resulting in higher energy dissipation, and electron transport chain impairment in both crops. Spectral reflectance indices suggested cold stress resulted in pigment differences between crops. The effect of priming was more pronounced in maize than in soybean with mostly a cumulatively negative effect. However, priming stabilized the electron trapping efficiency and upregulated the electron transfer system in maize, indicating an adaptive response. Overall, this comprehensive analysis provides insights into the complex physiological responses of maize and soybean to cold stress, emphasizing the need for further genotype-specific cold stress response and priming effect research.

First Report of Hemp Fusarium Wilt Caused by Fusarium oxysporum in Croatia.

Wilted hemp (Cannabis sativa L.) plants were observed in August 2019 in commercial fields around Osijek, Croatia. Plants and roots with disease symptoms were collected. The single-spored isolates produced septate cottony white to light pink aerial mycelium and purple undersurface on potato dextrose agar (PDA). Smooth and hyaline hyphae were branched and septate. Macroconidia were fusiform to sickle-shaped with foot-shaped basal cells, elongated apical cells and three to five septa. Sequencing of the internal transcribed spacer and the partial elongation factor 1-α gene identified the species as Fusarium oxysporum. Artificial infection fulfills Koch's postulates, producing plants which show stunted growth and wilt symptoms similar to those observed in the commercial fields. Control seedlings remained symptomless and healthy. To the best of our knowledge, this is the first report of hemp Fusarium wilt causing F. oxysporum in Croatia. Considering that F. oxysporum has been reported in main field crops in Croatia, the presence of this pathogen could cause economically significant hemp production decreases, especially in humid and cold springs and susceptible varieties.

First Report of Charcoal Rot Caused by Macrophomina phaseolina on Hemp in Croatia.

The production of industrial hemp (Cannabis sativa) has had a sharp increase in the past five years in Croatia (Mihelcic 2017). Production has been in constant increase, from 1560 ha in 2015 to 2476 ha in 2019 (PAAFRD). In August 2019, numerous (~1.5%) premature wilted hemp plants (cultivar Fibranova) were observed in commercial fields in Vladislavci (45.4646950° N, 18.5674770° E), around Osijek, Croatia. Diseased plants showed symptoms of chlorosis, rapidly wilting, necrosis and prematurely dying. The stalk of diseased plants was completely desiccated, while internal tissues were spongy and fluffy. Near the soil line, discoloration of the stalk with small spherical to oblong black microsclerotia was detected. Roots were necrotic with brown-gray areas. Twenty root and crown segments of the infected plants (2- to 3-mm long pieces) were surface sterilized with 2% NaOCl for 5 min, rinsed three times with sterile distilled water, and plated on potato dextrose agar (PDA, pH 6.2) media, containing 250 mg per liter of chloramphenicol to inhibit bacterial growth (Zveibil and Freeman 2005). The culture plates were incubated at 28 ± 2°C for 7 days in the dark and purified to obtain a pure culture that produces numerous, dark, hard, ovoidal-shaped sclerotia, averaging 140 x 52 µm (n=50). The single sclerotium isolate produced flat light to dark gray colonies with entire margins. Based on field symptoms, colony and microsclerotia morphology, the fungus was identified as Macrophomina phaseolina (Tassi) Goid (Marquez et al. 2021). Total DNA from the isolate was extracted with Extract-N-Amp Plant PCR Kit (Sigma-Aldrich Co., USA). To confirm morphological identification, part of the TEF 1-alpha gene region was amplified using EF1-728F (Carbone and Kohn 1999) and EF2 (O'Donell et al. 1998). The sequence of the isolate MP1 (212 bp - GenBank accession no. OQ389757), showed 100% nucleotide sequence identity to the reference sequence of M. phaseolina GenBank sequence MG434668 (Casano et al. 2018). Eighteen hemp plants (cv. Fibranova) were sown in six plastic pots (three hemp plants per pot) for the pathogenicity test. Ten-day-old M. phaseolina culture (isolate MP1) was used for inoculum preparation. Each pot of one-week-old plant was irrigated with 100 ml of a microsclerotia suspension (105 microsclerotia/ml)(Abied et al. 2018). Plants were held at 28°C and 70% relative humidity in a growth chamber (aralab, Fitoclima 10.000 HP) with a 16-hour photoperiod. Pots with control plants were irrigated with the same amount of sterile distilled water. Ten weeks after inoculation percentage of wilted plants was 77.78%. In the control variant all plants were healthy. M. phaseolina was reisolated from inoculated plants and morphologically identified. With the liberalization of the law, hemp production in Croatia is increasing, which could result in general disease problems and the disease caused by M. phaseolina. Charcoal rot will be expressed in years with dry and warm summers (Lodha and Mawar 2020), and relatively short, cool, rainy winters (Nevo et al. 2012), which has become common in the last decade in Croatia. To our knowledge, this is the first report of M. phaseolina on hemp in Croatia. The authors declare no conflict of interest.

Quantitative trait loci mapping of metal concentrations in leaves of the maize IBM population.

Characterizing concentrations of several beneficiary and toxic metals in maize leaves is of importance for ionomic studies and for silage production. The intermated B73 × Mo17 maize population (IBM) was evaluated for concentrations of eight metals (cadmium - Cd, copper - Cu, iron - Fe, potassium - K, magnesium - Mg, manganese - Mn, strontium - Sr and zinc - Zn) in ear-leaf to map quantitative trait loci (QTL) with 2161 molecular markers across the genome. QTL analysis revealed nine significant QTLs for concentrations of Cd, Cu, Fe, K, Mg and Sr combined over two environments. Median resolution for the QTL interval was less than 1 cM on a regular F2 map, which is a big improvement compared with the prior mapping (8 cM). The highest LOD scores of 15.52 and 15.31 were detected for K and Cd concentrations, respectively, explaining more than 20 percent of the phenotypic variance. No QTLs were found to be colocalized. QTL mapping in the IBM population did not confirm our earlier QTL results demonstrating considerable QTL ×genetic background interaction. The only exception is confirmation of the major QTL for Cd accumulation on chromosome 2. Our results could facilitate further genetic and physical mapping of genes for metal accumulation in maize.

Xanthium italicum, Xanthium strumarium and Arctium lappa as new hosts for Diaporthe helianthi.

Sunflower (Helianthus annuus) stem canker caused by Diaporthe helianthi is one of the most important sunflower diseases in Croatia. Until recently, sunflower was the only known host for D. helianthi. In our research carried out in the area of Eastern Croatia, isolates of Diaporthe/Phomospis were collected from Xanthium italicum, X. strumarium and Arctium lappa. Using morphological, cultural and molecular ITS rDNA data, isolates from these weeds were identified as D. helianthi. The following isolates were used in the pathogenicity test: one isolate originated from sunflower (Su5/04), three from X. italicum (Xa2, Xa3 and Xa5), two from X. strumarium (Xa9 and Xa12), one from Xanthium sp. (Xa13) and one from A. lappa (Ar3). According to the results, it was determined that isolate Xa5 (originated from X. italicum) was the most pathogenic to sunflower stems. The average length of the lesion was 11.3 cm. The lowest level of pathogenicity was found in Xa9 (isolated from X. strumarium). The length of the lesion was 0.1 cm.