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Flax (Linum usitatissimum L.) is a commercial oilseed crop in Canada. Globally flax is known for industrial oil and fiber. Flaxseed contains Omega 3 fatty acid, lignans like secoisolariciresinol diglucoside (SDG), flavonoids and polysaccharides which offer potential health benefits. Conventional flax cultivars are brown seeded and few mutant lines are yellow seeded. The darkness of seed colour depends on the presence of polymerized proanthocyanidins (PA; condensed tannins) in the seed coat. PAs are the product of the phenylpropanoid pathway. Previous genetic studies by Mittapalli and Rowland (2003) on G1186/94 showed the seed colour trait was governed by the homozygous recessive alleles at D locus and the same locus is closely linked to white or pink flower petals. To start with, single seeds of already developed stable recombinant inbred lines (RILs) (of F8:9 generation) from a cross of yellow seeded European recessive line (G1186/94) and brown seeded CDC Bethune (popular variety) were grown. In this study, seed colour phenotyping was done by measuring seed colour of each RIL in Red-Green-Blue (RGB) values. To understand the genetic basis of flax seed and flower colour, mapping with single sequence repeats (SSRs) and CAPS (Cleaved Amplified Polymorphic Sequences) markers were used. For the first time, a framework genetic linkage map was constructed from populations of CDC Bethune/ G1186/94 containing 19 linkage groups (LGs). LG 1 with four SSR markers was found to be linked with the seed colour locus D. During the fine-mapping, two SSR markers (LuM566 and Lu2351) were found to be linked with the seed colour trait. The D locus has been confined in a 2.8 cM region and the closest marker was LuM566 at a distance of 0.6 cM. This was observed to be a stable locus in two growth trials and in different environments with logarithm of odds (LOD) above 39 and more than 84 % of the trait expressed by the major locus in both trials. As there were no recombinants (off types) for flower colour in F8:9 plants i.e brown-seeded lines produced blue flowers and yellow-seeded lines produced white flowers, the same locus holds well for the flower colour trait. The marker associated with seed and flower colour in G1186/94 (European recessive yellow line) was identified and can be used in flax breeding. Additionally, an interesting putative candidate gene of potential significance was identified through genomics assisted gene search from the flax whole genome sequence database. The gene expression analyses showed lower expression of putative flavonoid 3’ hydroxylase (F3’H) (a gene involved in flavonoid biosynthesis pathway) in both seed coat and flower petal tissues of G1186/94 as compared to CDC Bethune. Therefore, this study represents the first report on genetic mapping based putative candidate gene finding for recessive yellow seed colour mutation in the D locus in flax.

Tikslas Vienuolikos rinktinių medžių klonų sėklinių palikuonių ir medžių išaugintų iš vietinių sėklų, kiekybinių ir kokybinių parametrų palyginimas. Objektas Želdiniai įveisti 1 ha plote 1980 metais Šakių u-jos Klevinių g-je pagal doc. J. Danusevičiaus projektą. Sodinimui plotas parinktas 140 kv. 6 skl. (1978 metų plynai iškirstoje degvieteje). Ploto dirvožeminė-¬tipologinė grupė - Nbl. Metodika Matavimai atlikti laikinuose nevienodo ploto tyrimo bareliuose, apribotuose eglių eilėmis. Visuose bareliuose išmatuoti visi ten augantys medžiai. Medžių aukštis ir aukštis iki žalių šakų matuotas 0,5 m tikslumu, skersmuo - 1,3 m aukštyje 1 cm tikslumu. Kiti parametrai buvo vertinami vizualiai. Stiebo tiesumas, šakų storis, šakų prisegimo kampas ir medžių būklė vertinti 5 balų sistema. Taip pat buvo vertinta medžių priklausomybė Krafto klasėms (I-V). Matavimų duomenys apdoroti Statistica programa. Rezultatai Nustatyta, kad 30 metų amžiuje pagal medžių aukštį geriausi yra 4 ir 8 šeimų medžiai (19,6 – 16,6% aukštesni už kontrolinius želdinius), blogiausi – 14 ir 9 šeimų medžiai. Didžiausią stiebų skersmenį turi 10 ir 4 šeimų medžiai (14,2 – 12,7% storesni už kontrolinius želdinius), o mažiausią skersmenį – 14 ir 7 šeimos medžiai. Beveik analogiškais rodikliais šios šeimos pasižymėjo ir 18 metų amžiuje. Pagal išlikimą ir sukauptą tūrį trys geriausios šeimos produkuoja 176 – 145 m3/ha ir tuo 1,9-1,6 karto lenkia vietinės kilmės medžius bei 2,8-2,3 karto - blogiausios 3 šeimos medžius... [toliau žr. visą tekstą] / Purpose To compare qualitative and quantitative data of eleven selected clones of descendant seed and trees, that were cultivated from local seed. Object Afforestation that was cultivated in 1 hectare in 1980 in Kleviniai forestry of State forest of Sakiai by the project of associate professor J. Danusevičius. Plantation was selected in the area in 140th block and 6th plot (in the area that was entirely cut down in 1978). Metods Measurements are made in temporary areas that are not of the size and are surrounded by rows of fir trees. All trees that grow in these areas were measured. Height of trees and height up to green knocks are measured by the precision of 0,5 meter, diameter in 1,3 meters high- by the precision of 1 centimeter. Other measurements were evaluated visually. Straightness of stem, thickness of knocks, angle between stem and knock and the condition of trees were evaluated using a system of 5 grades. Dependency to Kraft classes (I-V) was also evaluated. Data of measurements was processed by Statistica program. Results The study showed that in the age of 30 years the best are 4th and 8th clones' tree families (19,6 – 16,6% higher than controlled plans), the worst – 14th and 16th clones' tree families. 10th and 4th clones' tree families have the biggest diameter of stem (14,2 – 12,7% thicker then controlled plans), and the smallest diameter has 14th and ir 7th clones' tree families. These tree families had almost the same measurements also in the ages... [to full text]

Phosphoenolpyruvate (PEP) carboxylase (PEPC) is a tightly controlled enzyme situated at a pivotal branchpoint of plant C-metabolism. Two physically and kinetically distinct oligomeric classes of PEPC exist in the endosperm of developing castor oil seeds (COS). Class-1 PEPC is a typical homotetramer composed of 107-kDa plant-type PEPC (PTPC) subunits, whereas the 910-kDa Class-2 PEPC hetero-octameric complex arises from a tight interaction between Class-1 PEPC and distantly related 118-kDa bacterial-type PEPC (BTPC) subunits. BTPC functions as both a catalytic and regulatory subunit of the allosterically-desensitized Class-2 PEPC, which has been hypothesized to support massive PEP-flux to malate for leucoplast fatty acid synthesis. Previous studies established that BTPC: (i) subunits of COS Class-2 PEPC are subject to inhibitory phosphorylation in vivo, and (ii) at Ser425 and Ser451 within an intrinsically disordered region. This study focuses on characterization of the COS protein kinase (BTPC-K) that phosphorylates BTPC at Ser451. BTPC-K, having a native molecular mass of 63 kDa, was purified ~500-fold from developing COS endosperm. Its activity was absolutely dependent upon the presence of Ca2+ (Ka= 2.7 μM) and millimolar Mg2+. BTPC-K phosphorylated BTPC subunits of Class-2 PEPC strictly at Ser451 (Km= 1.1 μM), as well as histone type III-S (Km= 1.7 μM), but did not phosphorylate a BTPC S451D phosphomimetic mutant, native COS PTPC or sucrose synthase, or α-casein. BTPC-K displayed a broad pH-activity optima of pH 7.3, a Km for Mg2+-ATP of 6.6 μM, and marked inhibition by 3-P-glycerate and PEP. The possible control of BTPC-K by disulfide-dithiol interconversion was suggested by its rapid inactivation and subsequent reactivation when incubated with oxidized glutathione and then dithiothreitol. BTPC-K activity was insensitive to exogenous calmodulin, but potently inhibited by 100 µM trifluoperazine (a calmodulin antagonist). BTPC-K-mediated Ser451 phosphorylation of BTPC subunits of Class-2 PEPC inhibited BTPC activity by ~50% when assayed under suboptimal conditions (pH 7.3, 1 mM PEP with 10 mM L-malate). Overall the results of this study have led to the hypothesis that in vivo phosphorylation of COS BTPC at Ser451 is mediated by a dedicated calcium-dependent protein kinase (CDPK). / Thesis (Master, Biology) -- Queen's University, 2013-08-30 14:23:39.648

Magistrantūros studijų baigiamajame darbe pateikiami 2013 metų vasarinių kviečių veislių, sėtų po skirtingų priešsėlių, derlingumo ir sėklų kokybės rodiklių duomenys ir atliktas priešsėlių poveikio veislėms įvertinimas. Darbo objektas - vasariniai kviečiai 'Triso' ir 'Hamlet', sėti po skirtingų priešsėlių (vasarinių miežių ir vasarinių rapsų). Darbo metodai ir vieta. Tyrimai vykdyti 2013 m. Vilmos Žliobienės ūkyje, Antušavos kaime, Kėdainių rajone. Eksperimento įrengimo vietoje dirvožemis karbonatingas glėjiškas rudžemis (RDg4-k2). Buvo nustatytas grūdų derlingumas, derliaus struktūros komponentai (produktyvių stiebų skaičius, vidutinis grūdų skaičius varpoje, 1000 grūdų masė, varpos produktyvumas). Grūdų kokybės rodikliai (baltymų, šlapiojo glitimo, krakmolo kiekis, sedimentacijos rodiklis ir kiti) nustatyti AB :Linas agro" laboratorijoje. Darbo rezultatai. Esmingai didžiausias derlingumas (5,9 t ha-1) ir 1000 grūdų masė (43,84 g) nustatyti `Hamlet` veislės vasariniams kviečiams po vasarinių rapsų priešsėlio. Patikimai didžiausias produktyvių stiebų skaičius (432,8 vnt. m-2) ir vidutinis grūdų skaičius varpoje (34,8 vnt.) buvo `Triso` veislės augalų. Genotipo (veislės) savybės ir 1000 grūdų masė turėjo didesnę įtaką grūdų derlingumui ir esmingai padidino ‘Hamlet` veislės derlingumą. Esmingai didžiausias vasarinių kviečių grūdų baltymų (14,9 proc.) ir sedimentacijos rodiklis (71 proc.) nustatytas ‘Triso‘ veislės grūduose po vasarinių rapsų priešsėlio. Esmingai didžiausias... [toliau žr. visą tekstą] / In Master's thesis is a submitted spring wheat variety of 2013, which were sown under different preceding crops, yield and seed quality indicators in data and performed assessment of preceding crop varieties impact. The object. Spring wheat 'Triso' and 'Hamlet', which were sown after different preceding crops (spring barley spring oilseed rape). Working methods and location. Research was carried out in 2013 at Vilmos Žliobienės farm in the Antušavos village in the Kėdainių area. The soil is endohypogleyic cambisol (RDg4-k2) at installation site of the experiment. It was established grain yield, harvest structure components (the number of productive stems, average number of wheat-ear, 1000 grain weight, productivity of wheat-ear). Grain quality indicators (protein, wet gluten, starch content sedimentation rate and others) were determined at AB Linas Agro laboratory. Results. Essentially the highest grain yield (5.9 t ha-1) and 1000 grain weight (43.84 g) were obtained in the fields of `Hamlet` variety, which were sown after spring oilseed rape. Reliably largest number of productive tillers number (432.8 units. m-2) and the average number of grains per ear (34.8 pc) was the variety of `Triso` compared to the variety of `Hamlet`, which was investigated in the same influence of forecrop. But a variety of properties and weight of 1,000 grain had a greater impact on grain yield and substantially increased varieties of 'Hamlet `yield. Substantially the largest spring wheat grain... [to full text]

Canola (Brassica napus) contributes $19.3 billion dollars to the Canadian economy each year as a result of its oil- and protein-rich seeds. These economically important seed products are produced in highest concentration in the embryo. Embryo development is supported nutritionally and structurally by the maternal subregions, which include the inner (ISC) and outer distal seed coat (OSC), the chalazal seed coat (CZSC), and the chalazal proliferating tissue (CPT). Research on the maternal seed subregions is limited to the SC as a result of its accessibility; the embedded CZSC and CPT subregions have yet to be characterized in canola. Using light and transmission electron microscopy, I found the CZSC and CPT to be anatomically distinct and experience profound changes throughout seed development. To understand these changes at the RNA level, laser microdissection and RNA sequencing were used to profile these subregions spatially and temporally from the ovule to mature green stage of seed development. Employing vigorous bioinformatics analyses, I found that the maternal subregions are transcriptomically distinct and possess unique RNA populations. From here I began to elucidate the biological processes operating within the maternal subregions. As a whole, the maternal subregions appear to have a critical role in transporting nutrients to the filial subregions as well as in coping with oxidative stress produced during these energy-rich processes. Additionally, using CanEnrich, I was able to generate predictive transcriptional circuits regulating the biological processes occurring within the maternal seed. This research has produced the most comprehensive dataset on the canola seed to date and will provide a valuable resource for research on seed development as well as seed improvement. / October 2016

Due to the immense utilization in food and industry, there is enormous commercial and scientific interest to manipulate canola (Brassica napus L.) seed oil. Seed oil accretions are influenced by genes involved in embryo and seed development. FUSCA3 (FUS3) and LEAFY COTYLEDON1 (LEC1) are well-known transcription factors involved during seed and embryo development. The main objective of this project was to evaluate the role of these genes during seed storage deposition and microspore-derived embryogenesis in B. napus. For this purpose, six BnLEC1 transgenic lines and three BnFUS3 TILLING mutant lines were generated. The over expression of BnLEC1 significantly increased the seed oil content, while the down regulation of BnLEC1 or mutation of BnFUS3 reduced the level of seed oil. Experimental alterations of BnLEC1 and BnFUS3 triggered transcriptional modifications in enzymes taking part in sucrose transport and metabolism, glycolysis, and fatty acid (FA) biosynthesis. These changes are suggestive of a greater carbon pool to FA biosynthesis in tissues over-expressing BnLEC1, and a reduced carbon flux available for the synthesis of FA in BnLEC1 down regulators and BnFUS3 tilling mutants. While the elevated oil content induced by BnLEC1 was not accompanied by alterations in FA composition, oil nutritional value, or glucosinolate (GLS) levels, suppression of BnLEC1 reduced seed oil accumulation and raised levels of GLS, possibly through the transcriptional regulation of BnST5a (Sulphotransferase5a), the last GLS biosynthetic enzyme. BnFUS3 tilling mutant seeds had increased levels of linoleic acid, possibly due to the reduced expression of ω-3 FA DESATURASE (FAD3). The effects of altered expression of BnLEC1 and BnFUS3 were also assessed during microspore-derived embryogenesis. Substantial structural abnormalities, accompanied by changes in transcript levels of several embryo marker genes were observed in embryos in which the expression of BnLEC1 or BnFUS3 was altered. The changes in oil level and FA profiles observed in the transformed microspore-derived embryos followed a similar trend to that described in seeds. Collectively, these observations suggest that manipulation of BnLEC1 and BnFUS3 can be employed as a tool to enhance seed oil production and quality in B. napus. / February 2017