Basil is one of the most widespread aromatic and medicinal plants, which is often grown in drought- and salinity-prone regions. Often co-occurrence of drought and salinity stresses in agroecosystems ...and similarities of symptoms which they cause on plants complicates the differentiation among them. Development of automated phenotyping techniques with integrative and simultaneous quantification of multiple morphological and physiological traits enables early detection and quantification of different stresses on a whole plant basis. In this study, we have used different phenotyping techniques including chlorophyll fluorescence imaging, multispectral imaging, and 3D multispectral scanning, aiming to quantify changes in basil phenotypic traits under early and prolonged drought and salinity stress and to determine traits which could differentiate among drought and salinity stressed basil plants.
"Genovese" was grown in a growth chamber under well-watered control 45-50% volumetric water content (VWC), moderate salinity stress (100 mM NaCl), severe salinity stress (200 mM NaCl), moderate drought stress (25-30% VWC), and severe drought stress (15-20% VWC). Phenotypic traits were measured for 3 weeks in 7-day intervals. Automated phenotyping techniques were able to detect basil responses to early and prolonged salinity and drought stress. In addition, several phenotypic traits were able to differentiate among salinity and drought. At early stages, low anthocyanin index (ARI), chlorophyll index (CHI), and hue (HUE
), and higher reflectance in red (R
), reflectance in green (R
), and leaf inclination (LINC) indicated drought stress. At later stress stages, maximum fluorescence (F
), HUE
, normalized difference vegetation index (NDVI), and LINC contribute the most to the differentiation among drought and non-stressed as well as among drought and salinity stressed plants. ARI and electron transport rate (ETR) were best for differentiation of salinity stressed plants from non-stressed plants both at early and prolonged stress.
Micronutrient malnutrition is one of the main public health problems in many parts of the world. This problem raises the attention of all valuable sources of micronutrients for the human diet, such ...as common bean (
L.). In this research, a panel of 174 accessions representing Croatian common bean landraces was phenotyped for seed content of eight nutrients (N, P, K, Ca, Mg, Fe, Zn, and Mn), and genotyped using 6,311 high-quality DArTseq-derived SNP markers. A genome-wide association study (GWAS) was then performed to identify new genetic sources for improving seed mineral content. Twenty-two quantitative trait nucleotides (QTN) associated with seed nitrogen content were discovered on chromosomes Pv01, Pv02, Pv03, Pv05, Pv07, Pv08, and Pv10. Five QTNs were associated with seed phosphorus content, four on chromosome Pv07, and one on Pv08. A single significant QTN was found for seed calcium content on chromosome Pv09 and for seed magnesium content on Pv08. Finally, two QTNs associated with seed zinc content were identified on Pv06 while no QTNs were found to be associated with seed potassium, iron, or manganese content. Our results demonstrate the utility of GWAS for understanding the genetic architecture of seed nutritional traits in common bean and have utility for future enrichment of seed with macro- and micronutrients through genomics-assisted breeding.
In Croatia, the majority of the common bean production is based on local landraces, grown by small-scale farmers in low input production systems. Landraces are adapted to the specific growing ...conditions and agro-environments and show a great morphological diversity. These local landraces are in danger of genetic erosion caused by complex socio-economic changes in rural communities. The low profitability of farms and their small size, the advanced age of farmers and the replacement of traditional landraces with modern bean cultivars and/or other more profitable crops have been identified as the major factors affecting genetic erosion. Three hundred accessions belonging to most widely used landraces were evaluated by phaseolin genotyping and microsatellite marker analysis. A total of 183 different multi-locus genotypes in the panel of 300 accessions were revealed using 26 microsatellite markers. Out of 183 accessions, 27.32% were of Mesoamerican origin, 68.31% of Andean, while 4.37% of accessions represented putative hybrids between gene pools. Accessions of Andean origin were further classified into phaseolin type II ("H" or "C") and III ("T"), the latter being more frequent. A model-based cluster analysis based on microsatellite markers revealed the presence of three clusters in congruence with the results of phaseolin type analysis.
Crop production might suffer severe economic losses due to insufficient fertiliser availability. Specific signs of nutrient shortage influence plant morphology and physiology. This study pioneers the ...non-destructive tracking and characterization of nutrient deficiency symptoms in common beans using multispectral and chlorophyll fluorescence imaging, offering novel insights into the dynamic responses of plant morphology and physiology to specific nutrient shortages. Plants were cultivated in nutrient solutions with and without nitrogen (N), phosphorus (P), potassium (K), magnesium (Mg), iron (Fe) and control solution. Measurements were taken every three days for 12 days (MT1-MT4) of growth. K and N deficit plants had the earliest symptoms and most noticeable changes, whereas Fe deficiency plants had the slowest and least noticeable symptoms. Except for Fe, the most responsive chlorophyll fluorescence parameter was electron transport rate, which was reduced in plants from all nutrient deprivation treatments compared to control. All nutrient deficit treatments reduced leaf area at MT2, which was the most affected morphological parameter. The green leaf index, reflection in blue, and specific green were the most affected multispectral traits by nutritional deprivation. These findings demonstrate that plant nutrient deficit can be recognized and tracked non-destructively utilizing multispectral and chlorophyll fluorescence analysis. Overall, our work not only sheds light on the dynamics of nutrient deficiency in common bean plants but also offers practical implications for improving crop management strategies using non-destructive digital technology.
Winter savory (Satureja montana L.) is an important medicinal, aromatic, and honey plant. In Croatia, it is widely distributed along the Adriatic coast, where it is frequently exposed to droughts. ...First, the winter savory natural population with the highest germination across different drought treatments after hydropriming was selected. Nine hundred seeds from each of the three natural populations (P1, P2, and P3) were hydroprimed (dH2O) for 48 h. The seeds were then germinated in drought treatments with different concentrations of polyethylene glycol (PEG 6000) (−0, −0.2, −0.4, −0.8, −1.2, −1.6, −2, −2.5, −3.0 MPa). Since P1 showed the best results in germination parameters, it was used for the second phase of the experiment, where the effect of hormonal priming (100 and 400 ppm GA3, 48 h in the dark) and osmopriming (0.2% and 0.6% w/v KNO3, 72 h in the dark) on seed germination and seedling morphological parameters of the selected winter savory population under drought stress conditions (−0.8 and −2.5 MPa) was evaluated. Although winter savory grows in dry areas, this study showed that extremely dry conditions (−3.0 and −2.5 MPa) negatively affected seed germination, but this effect can be mitigated by priming treatments, especially with the hormonal priming (GA3 400 ppm).
Drought is a significant constraint in bean production. In this study, we used high-throughput phenotyping methods (chlorophyll fluorescence imaging, multispectral imaging, 3D multispectral scanning) ...to monitor the development of drought-induced morphological and physiological symptoms at an early stage of development of the common bean. This study aimed to select the plant phenotypic traits which were most sensitive to drought. Plants were grown in an irrigated control (C) and under three drought treatments: D70, D50, and D30 (irrigated with 70, 50, and 30 mL distilled water, respectively). Measurements were performed on five consecutive days, starting on the first day after the onset of treatments (1 DAT-5 DAT), with an additional measurement taken on the eighth day (8 DAT) after the onset of treatments. Earliest detected changes were found at 3 DAT when compared to the control. D30 caused a decrease in leaf area index (of 40%), total leaf area (28%), reflectance in specific green (13%), saturation (9%), and green leaf index (9%), and an increase in the anthocyanin index (23%) and reflectance in blue (7%). The selected phenotypic traits could be used to monitor drought stress and to screen for tolerant genotypes in breeding programs.
Landraces represent valuable genetic resources for breeding programmes to produce high-yielding varieties adapted to stressful environmental conditions. Although the common bean (
L.) is an ...economically important food legume for direct human consumption worldwide, common bean production in Croatia is based almost exclusively on landraces and there is no common bean breeding program. Information on phaseolin type and results of population structure and genetic diversity obtained by analysis of SSR and SNP markers, in combination with the morphological characterization of 174 accessions of 10 common bean landraces (morphotypes), enabled thorough classification of accessions. The accessions were classified into phaseolin type H1 ("S") of Mesoamerican origin and phaseolin types H2 ("H" or "C") and H3 ("T") of Andean origin. By applying distance- and model-based clustering methods to SSR markers, the accessions were classified into two clusters at
= 2 separating the accessions according to the centres of origin, while at
= 3, the accessions of Andean origin were further classified into two clusters of accessions that differed in phaseolin type (H2 and H3). Using SNP markers, model-based analysis of population structure was performed, the results of which were consistent with those of SSR markers. In addition, 122 accessions were assigned to 14 newly formed true-type morphogenetic groups derived from three different domestication events: (1) Mesoamerican (H1A) ("Biser", "Kukuruzar", "Tetovac", "Trešnjevac"), (2) Andean-indeterminate type (H2B1) ("Dan noć", "Sivi", "Puter", "Sivi prošarani", "Trešnjevac") and (3) Andean-determinate type (H3B2) ("Bijeli", "Dan noć", "Puter", "Trešnjevac", "Zelenčec"). The rest of the accessions could represent putative hybrids between morphogenetic groups. The differences between the true-type groups of accessions were further analysed based on nine quantitative traits, and the subsets of traits that best distinguish among centres of origin (A: Mesoamerican, B: Andean) and genetic groups (H1A, H2B1, H3B2) were proposed.
Sweet pepper (Capsicum annuum L.) is one of the most important vegetable crops in the world because of the nutritional value of its fruits and its economic importance. Calcium (Ca) improves the ...quality of sweet pepper fruits, and the application of calcite nanoparticles in agricultural practice has a positive effect on the morphological, physiological, and physicochemical properties of the whole plant. The objectives of this study were to investigate the effect of commercial calcite nanoparticles on yield, chemical, physical, morphological, and multispectral properties of sweet pepper fruits using a combination of conventional and novel image-based nondestructive methods of fruit quality analysis. In the field trial, two sweet pepper cultivars, i.e., Šorokšari and Kurtovska kapija, were treated with commercial calcite nanoparticles (at a concentration of 3% and 5%, calcite-based foliar fertilizer (positive control), and water (negative control) three times during vegetation). Sweet pepper fruits were harvested at the time of technological and physiological maturity. Significant differences were observed between pepper cultivars as well as between harvests times. In general, application of calcite nanoparticles reduced yield and increased fruit firmness. However, different effects of calcite nanoparticles were observed on almost all properties depending on the cultivar. In Šorokšari, calcite nanoparticles and calcite-based foliar fertilizers significantly increased N, P, K, Mg, Fe, Zn, Mn, and Cu at technological maturity, as well as P, Ca, Mg, Fe, Zn, Mn, Cu, and N at physiological maturity. However, in Kurtovska kapija, the treatments increased only Ca at technological maturity and only P at physiological maturity. The effect of treatments on fruit morphological properties was observed only at the second harvest. In Šorokšari, calcite nanoparticles (3% and 5%) increased the fruit length, minimal circle area, and minimal circle radius, and it decreased the fruit width and convex hull compared to the positive and negative controls, respectively. In Kurtovska kapija, calcite nanoparticles increased the fruit width and convex hull compared to the controls. At physiological maturity, lower anthocyanin and chlorophyll indices were found in Kurtovska kapija in both treatments with calcite nanoparticles, while in Šorokšari, the opposite effects were observed.
Common bean (Phaseolus vulgaris L.) exhibits a wide range of seed coat colors and this morphological trait is widely used in cultivar identification and assessment of diversity within this species. ...With an advancement in technology and informatics, new methods of assessing seed color are emerging in addition to traditionally used visual observation. Due to a great variety of color measuring techniques, the evaluation of the agreement between methods is needed prior to using the methods interchangeably. Seed coat color in terms of CIE L*a*b* color coordinates of 100 common bean accessions belonging to five mono-colored landraces was assessed using two methods, colorimeter and Computer vision. The percentage difference between the two methods across all samples for L* color coordinate was 5.81%, for a* color coordinate 23.32% and for b* color coordinate 44.44%. According to Bland-Altman difference plot there is a considerable lack of agreement between the two methods. However, using stepwise discriminant analysis revealed that colorimeter method correctly classified 97% of accessions into their respective landrace, while the classification success of the Computer vision was 99%. Keywords: colorimetry, common bean, Computer vision, landrace Grah (Phaseolus vulgaris L.) pokazuje veliki raspon boja sjemene ljuske te je koristenje ove morfoloske karakteristike uobicajeno u identifikaciji kultivara i procjeni raznolikosti unutar vrste. Napretkom tehnologije i informatike pojavljuju se nove metode odredivanja boje sjemena uz klasicnu metodu vizualnog odredivanja. Zbog velikog broja tehnika odredivanja boja, potrebna je ocjena usuglasenosti izmedu metoda prije njihovog kombiniranja ili naizmjenicnog koristenja. Boja sjemena 100 primki pet jednobojnih tradicijskih kultivara graha opisana CIE L*a*b* koordinatama procijenjena je koristeci dvije metode, kolorimetar i racunalni vid. Postotna razlika izmedu dvije metode kroz sve uzorke za koordinatu L* iznosila je 5.81%, za koordinatu a* 23.32%, a za koordinatu b* 44.44%. Bland-Altmanov graf razlike pokazuje da postoji znacajan nedostatak usuglasenosti izmedu dvije metode. Koristeci postupnu diskriminantnu analizu uspjesno je klasificirano 97% primki na temelju podataka dobivenih kolorimetrom dok je klasifikacijski uspjeh racunalnog vida bio 99%. Kljucne rijeci: grah, kolorimetrija, racunalni vid, tradicijski kultivari
Common bean is one of the most cultivated and consumed grain legumes worldwide, showing a high level of genetic diversity. Here is presented a detailed review of development and mapping of simple ...sequence repeats (SSRs, microsatellite markers) in the common bean. In the last 25 years, common bean has been the subject of numerous genetic studies, in which the identification and use of SSRs were conducted, and lead to the development of genetic maps. First genetic maps of common bean have been developed in the 1990s and were based on different molecular markers, and included domestication genes and important agronomic traits. Later, SSRs allowed the genetic mapping of more narrow crosses that are often of interest in plant breeding. Most genetic maps have been correlated with the core map established in the recombinant inbred population BAT93 x Jalo EEP558, and includes different markers, RFLP (restriction fragment length polymorphism), RAPD (random amplified polymorphic DNA), AFLP (amplified fragment length polymorphism), and SSRs in particular. More than 2,000 SSR markers are available for the common bean and they are an important tool to evaluate the genetic diversity of common bean landraces. SSRs are also useful to evaluate intra-specific diversity within the genus Phaseolus. Keywords: common bean, genetic diversity, genetic maps, microsatellite markers Grah je jedna od najvise uzgajanih i najvise koristenih zrnatih mahunarki u svijetu koja ima visoku razinu genetske raznolikosti. U ovom radu je prikazan detaljan pregled razvoja i kartiranja mikrosatelitnih biljega (SSR, ponavljajuce jednostavne sekvence) kod graha. U posljednjih 25 godina, grah je bio predmetom brojnih genetskih istrazivanja u kojima je provedena identifikacija i upotreba SSR-ova sto je dovelo do razvoja genetskih karata. Prve genetske karte graha su razvijene u 1990-ima i bile su bazirane na razlicitim molekularnim biljezima, a ukljucivale su gene koji su povezani uz udomacenje i vazna agronomska svojstva. Kasnije su SSR-ovi omogucili genetsko kartiranje krizanaca koji su cesto od interesa u oplemenjivanju bilja. Vecina genetskih karata povezana je s osnovnom genetskom kartom uspostavljenom na rekombinantnoj inbred liniji BAT93 x Jalo EEP558, koja ukljucuje razlicite biljege kao sto su RFLP (polimorfizam duzine restrikcijskih ulomaka), RAPD (nasumicno umnozena polimorfna DNA), AFLP (polimorfizam duzine umnozenih ulomaka) te SSR-ove. Vise od 2 000 SSR biljega je dostupno za grah i oni su vazan alat za procjenu genetske raznolikosti tradicijskih kultivara graha. SSR-ovi su takoder korisni za procjenu unutarvrsne raznolikosti unutar roda Phaseolus. Kljucne rijeci: grah, genetska raznolikost, genetske karte, mikrosatelitni biljezi
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