Spatial and Temporal Assessment of Brassica napus L. Maintaining Genetic Diversity and Gene Flow Potential: An Empirical Evaluation

Unpredicted persistence of all forms of B. napus present in the agro-ecosystem is the most common consequence of preservation and self-recruitment of seeds originating from soil seed bank. In nature, spontaneous intra- and inter-specific hybridization of B. napus is possible with sexually compatible species from the Brassicaceae family. The aim of this chapter is (a) to identify the distribution pattern and population dynamics of volunteers and feral populations along statistical regions in Slovenia; (b) to assess the global diversity of naturally appearing B. napus plants; (c) to evaluate the genetic differentiation between volunteers and feral populations; (d) to obtain the spatial and temporal distribution of spontaneous pollination potential and estimation of gene flow conservation; (e) to find the empirically assigned out-crossing rate of B. napus under a fragmented landscape structure, during 4-year monitoring; and (f) to observe that ecologically, evolutionary, and agronomically oriented studies could be conducted at the DNA level using short sequence repeat (SSR) markers. In total, we collected 261 samples of volunteer and feral populations. Our results showed that alleles from both volunteer and feral populations were distributed in three genetic clusters with relatively similar levels of diversity. Naturally occurring out-crossing rate is 13.71%. The global Mantel correlation coefficient of genetic and spatial relatedness between genotypes is 0.044

DETECTION OF RECESSIVE MUTATIONS (CVM, BLAD AND RED FACTOR) INHOLSTEIN BULLS IN SLOVENIA

Detection of recessive mutations that causes complex vertebral malformation (CVM) and bovine leukocyte adhesion defi ciency (BLAD) in Holstein cattle is especially required for bulls, which are used for artifi cial insemination (A.I.); these enable elimination of carriers from the A.I. programs and therefore prevent transmission of unwanted mutations to a large number of offspring. Some breeders are also interested in the identifi cation of carriers of recessive allele for red and white coat colour (Red factor). Here, we performed genetic tests for detection of mutations associated with CVM, BLAD and Red factor using methods previously reported or modifi ed methods. Analysis of Holstein bulls, which were recommended for A.I in Slovenia in the years 2007 and 2008, revealed four (10 %) carriers of CVM, and two (5.4 %) carriers of red gene, while all bulls were non-carriers of BLAD

DOLOČANJE RECESIVNIH MUTACIJ (CVM, BLAD IN ALEL ZA RDEČO BARVO) PRI BIKIH ČRNO-BELE PASME V SLOVENIJI

Detection of recessive mutations that causes complex vertebral malformation (CVM) and bovine leukocyte adhesion defi ciency (BLAD) in Holstein cattle is especially required for bulls, which are used for artifi cial insemination (A.I.); these enable elimination of carriers from the A.I. programs and therefore prevent transmission of unwanted mutations to a large number of offspring. Some breeders are also interested in the identifi cation of carriers of recessive allele for red and white coat colour (Red factor). Here, we performed genetic tests for detection of mutations associated with CVM, BLAD and Red factor using methods previously reported or modifi ed methods. Analysis of Holstein bulls, which were recommended for A.I in Slovenia in the years 2007 and 2008, revealed four (10 %) carriers of CVM, and two (5.4 %) carriers of red gene, while all bulls were non-carriers of BLAD.Pri bikih, ki so namenjeni za osemenjevanje, je zaželena detekcija mutacij povezanih s CVM (prirojena kompleksna vretenčna anomalija) in BLAD sindromom (odsotnost sposobnosti obrambe levkocitov), ker s pravočasnim izločanjem nosilcev, lahko preprečimo prenos nezaželenih mutacij na veliko število potomcev. V interesu nekaterih rejcev je tudi identifi kacija nosilcev recesivnega alela za rdeče-belo barvo dlake (Red factor). V tem delu smo izvedli detekcijo mutacij povezanih s CVM in BLAD oz. rdečo-belo barvo dlake (Red factor) z genetskimi testi po že znanih metodah oz. s spremenjenimi metodami. Rezultati analize vzorcev plemenskih bikov črno-bele pasme, ki so bili priporočeni za osemenjevanje v Sloveniji v letih 2007 in 2008 so bili naslednji: štirje biki (10 %) so nosilci mutacije, ki povzroča CVM, dva bika (5,4 %) sta nosilca alela za rdečo barvo, medtem ko so vsi biki prosti mutacije, ki povzroča BLAD

Uporaba DNA markerjev za genetsko diferenciacijo navadnega (Avena sativa L.) in golega ovsa (Avena nuda L.)

The usefulness of genetic identification of varieties for seed quality analyses becomes important, when we suspect the presence of another variety, species or even genus, based on morphological seed traits in the purity analysis and germination test, With genetic analyses, the doubt about the authenticity of the naked oats (Avena nuda L.) variety \u27Kamil\u27 was successfully solved. There were atypical seeds with chaff among the samples of this variety, so it was not possible to confirm with certainty, whether these were seeds of the same species/variety or impurities of common oats (Avena sativa L.). Based on the seed phenotype of the \u27Kamil\u27 variety (two independent samples 284 and 285), four sub-samples were prepared (284AN, 284AS, 285AN, 285AS); AN label was the sub-sample with naked seeds and AS label was the sub-sample with chaffed seeds. In addition, another variety (\u27Noni\u27) and four accessions of common oats (ACC378, ACC379, ACC380, ACC381) from the Slovene Plant Gene Bank were included as common oat standards. A total of 36 plant samples of Avena sp. were included in the genetic differentiation. Using a set of six highly informative SSR markers and the results of diversity parameters at individual loci and paired genetic comparisons, we were able to confirm that the analysed sub-samples belonged to the same variety, i.e. \u27Kamil\u27. In addition, despite the morphological similarity, the chaffed seeds of naked oats variety ‘Kamil’ were sufficiently (genetically) different from the analysed variety and/or accessions of common oats.Uporabnost genetske identifikacije sort se kaže pri analizah kakovosti semena, ko se na osnovi morfoloških znakov semena pri analizi čistote in/ali klic v postopku kalivosti pojavi sum na drugo sorto, vrsto ali celo rod. Z aplikativno uporabo genetskih analiz smo uspešno rešili dvom o pristnosti sorte \u27Kamil\u27 vrste golega ovsa (Avena nuda L.). Med semenom omenjene sorte je bilo namreč prisotno tudi seme s plevo, a se ni dalo z gotovostjo potrditi ali gre za seme iste vrste s plevo ali za primes semen navadnega ovsa (Avena sativa L.). Na podlagi fenotipa semena sorte \u27Kamil\u27 (dva neodvisna vzorca 284 in 285) so bili pripravljeni štirje pod-vzorci (284AN, 284AS, 285AN, 285AS), kjer AN pomeni golce, AS pa plevence. Kot standardi navadnega ovsa so bili v genetsko identifikacijo vključene še sorta navadnega ovsa \u27Noni\u27 in štiri akcesije navadnega ovsa (ACC378, ACC379, ACC380, ACC381) iz Slovenske rastlinske genske banke. Skupno je bilo v genotipizacijo vključenih 36 vzorcev rastlin rodu Avena sp. Z uporabo seta šestih visoko informativnih SSR markerjev in rezultatov analize parametrov raznolikosti na posameznih lokusih ter parnih genetskih primerjav smo lahko potrdili, da so analizirani pod-vzorci pripadali isti vrsti in sorti, t.j. golemu ovsu \u27Kamil\u27. Poleg tega so bili kljub morfološki podobnosti plevenci sorte Kamil‘ genetsko dovolj različni od analizirane sorte in/ali akcesij navadnega ovsa

Macedonian Genebank: Seed Protein Content of Wild Red Clover (Trifolium pratense L.) Accessions

During this study, the content of crude protein content in seeds of 23 wild red clover (Trifolium pratense L.) populations, collected in East part of Macedonia, district Probistip, was assessed. On the basis of the average crude protein content, the collected accessions were divided into 3 groups with the crude protein content ≥ 15.0% (MKD01527, MKD01530, MKD01531, MKD01539, MKD01540, MKD01542, MKD01543); ≥ 16.0% (MKD01525, MKD01528, MKD01529, MKD0538, MKD01541, MKD01553, MKD01554, MKD01560) and ≥ 17.0% (MKD01536, MKD01537, MKD01544). The highest crude protein content of 17.3% was determined for population MKD01535 and the lowest (14.4 %) for MKD01526. Information on protein content, as an indicator of wild red clover quality, could be used as a selection tool for identifying accessions to be included in red clover breeding programs

Macedonian Genebank: Seed Protein Content of Wild Red Clover (Trifolium pratense L.) Accessions

During this study, the content of crude protein content in seeds of 23 wild red clover (Trifolium pratense L.) populations, collected in East part of Macedonia, district Probistip, was assessed. On the basis of the average crude protein content, the collected accessions were divided into 3 groups with the crude protein content ≥ 15.0% (MKD01527, MKD01530, MKD01531, MKD01539, MKD01540, MKD01542, MKD01543); ≥ 16.0% (MKD01525, MKD01528, MKD01529, MKD0538, MKD01541, MKD01553, MKD01554, MKD01560) and ≥ 17.0% (MKD01536, MKD01537, MKD01544). The highest crude protein content of 17.3% was determined for population MKD01535 and the lowest (14.4 %) for MKD01526. Information on protein content, as an indicator of wild red clover quality, could be used as a selection tool for identifying accessions to be included in red clover breeding programs

Agronomic Evaluation and Utilization of Red Clover (Trifolium pratense L.) Germplasm

Germplasm collection as a source of variability and genetic diversity must be evaluated for an efficient management and effective utilization. This research was aimed to evaluate the agronomic value of red clover germplasm collection, to group the cultivars and population according to their morpho-agronomical traits, and to select valuable materials for future breeding programs. Thirty red clover cultivars and populations of different geographical origin were included in the study. Investigation was carried out at the experimental field of the Agricultural Institute Osijek in Croatia. The field trial was arranged in a randomized block design with four replications. Each plot included twenty spaced plants (50 x 50 cm) of each cultivar/population. During two consecutive years (2006, 2007) the following data were collected: yields of green mass, dry matter and seed (g/plant), plant height (cm), dry matter content (%), persistence (%), flowering time. The analysis of variance was carried out for all the traits, with the exception of seed yield and flowering time (which were not recorded as replicated data), using PROC GLM of SAS 9.1. Differences among cultivars/populations were highly significant for all analyzed traits. Hierarchical cluster analysis (Ward method) based on the seven morpho-agronomic traits allowed the identification of six groups of red clover cultivars/populations. This research shows that there is a great genetic variability in the investigated germplasm collection. The most promising materials were selected to form a new breeding gene pool that could be helpful for the improvement of our red clover breeding programme

Ultrasound influence on coleoptile length at Poaceae seedlings as valuable criteria in prebreeding and breeding processes

The study includes research on the effect of ultrasound on the ability of seed germination and coleoptile development of cereal landraces (fam. Poaceae): oat (Avena sativa L.) brodski, rye (Secale cereale L.) rakotinska, Triticale svetinikolsko and soft wheat (Triticum aestivum L.) govrlevska. The experiment with ultrasound was carried out at frequency of 30-40 kHz on the thermostatic ultrasonic bath. Seeds were sonicated at a constant temperature (25ºC) for 15 min. Ultrasound effect is reflected in the length of the coleoptyl and mesocotyl, although in a good deal is genetically predetermined. Coleoptile length was the longest in Triticale (3.3-5.5 cm) and approximately the same lengths are evident in rye (3.0-6.1 cm). Concerning the mesocotyl the longest one (5.525 cm ± 0.697) is recorded at rye, even significantly longer than control of oat and Triticale. Longer coleoptile of sonicated seeds indicates faster seedling development, good water supply and rapid development and emergence of first leaves. Correlation coefficient showed very high (0.821) and high R2 (67,472%) dependence among variables both, in length of coleoptiles (as dependent variable) and mesocotyl (as independent) with low standard error (0.225). With the simple use of ultrasound the germination period could be shorten, water supply more efficient, the sowing periods will be shorten and good yields even under conditions of climate change with increased temperatures could be achieved

Drought Stress Response in Agricultural Plants: A Case Study of Common Bean (<em>Phaseolus vulgaris</em> L.)

Drought is one of the major threats to common bean (Phaseolus vulgaris L.), affecting its growth and productivity and, thus, contributing to considerable losses in yield in many regions worldwide. The development of varieties tolerant of drought stress has, therefore, become one of the primary goals in many common bean breeding programs. Plants have developed various mechanisms of their adaptation, to a greater or lesser extent, to drought. These are expressed, on the molecular level, by changes of gene expression and of protein content, together with responses at physiological and morphological levels. The response of common bean to drought is still not sufficiently well characterized due to its genetic complexity and its diverse, often ambiguous, phenotypic effects. Understanding these mechanisms is thus of fundamental importance for developing varieties that are better adapted to such stress conditions. In this chapter, we present research that provides an insight into the morpho-physiological adaptation and its underlying molecular changes in common bean plants exposed to drought. We include our contribution to establishing the basis for breeding of common bean with greater tolerance to this abiotic stress that uses molecular markers and identification of quantitative trait loci (QTLs)