Quality of DNA for PCR extracted from sunflower with different methods

Studying of plants DNA with PCR method plays an important role in the agrarian activity. Preceding stage of DNA studying is its extraction from plant material. We studied quality of DNA extracted by the different methods from seeds, seedlings, and green leaves of sunflower. We used an inbred sunflower line as a research object. For analysis we used embryos, dry and presoaked during 24 h seeds, roots, 7-day seedlings, green leaves. DNA was extracted using five methods: 1 – a standard method of extraction (1% CTAB), 2 – a modified method (2% CTAB), 3–4 – extraction with sets (Diamond DNA Plant Kit D, Russia; Lumiprobe, Germany), 5 – extraction from green leaves (2% CTAB with absorbent carbon). Amplification was conducted in thermocycler S1000тм (BioRad, USA). Spectrophotometery was done at scanning spectrophotometer LEKI SS2110UV (Russia). Analyzing all used methods we concluded they allow extracting of DNA from dry seeds, seedlings and green leaves of sunflower with sufficient reliability and repeatability, it is proved by PCR results. We couldn’t extract DNA from roots and presoaked seeds with the modified method (2% CTAB). The most economically profitable is first method (1% CTAB). Due to the results of spectrophotometery, the highest level of DNA clearance can be reached with method 3 – extraction by a set Diamond DNA Plant Kit D from seedlings and method 5 – extraction by 2% СТАВ with absorbent carbon from green leaves. The method 3 is more preferable by time necessary for DNA extraction.

Organization Features of the Mitochondrial Genome of Sunflower (Helianthus annuus L.) with ANN2-Type Male-Sterile Cytoplasm

This study provides insights into the flexibility of the mitochondrial genome in sunflower (Helianthus annuus L.) as well as into the causes of ANN2-type cytoplasmic male sterility (CMS). De novo assembly of the mitochondrial genome of male-sterile HA89(ANN2) sunflower line was performed using high-throughput sequencing technologies. Analysis of CMS ANN2 mitochondrial DNA sequence revealed the following reorganization events: twelve rearrangements, seven insertions, and nine deletions. Comparisons of coding sequences from the male-sterile line with the male-fertile line identified a deletion of orf777 and seven new transcriptionally active open reading frames (ORFs): orf324, orf327, orf345, orf558, orf891, orf933, orf1197. Three of these ORFs represent chimeric genes involving atp6 (orf1197), cox2 (orf558), and nad6 (orf891). In addition, orf558, orf891, orf1197, as well as orf933, encode proteins containing membrane domain(s), making them the most likely candidate genes for CMS development in ANN2. Although the investigated CMS phenotype may be caused by simultaneous action of several candidate genes, we assume that orf1197 plays a major role in developing male sterility in ANN2. Comparative analysis of mitogenome organization in sunflower lines representing different CMS sources also allowed identification of reorganization hot spots in the mitochondrial genome of sunflower.

Dormancy in sunflower line A-3: the role of the pericarp

Sunflower (Helianthus annuus L.) can often display seed dormancy, which causes a delay for immediate sowing. The final degree of “whole seed” dormancy is determined by the contributions of the tissues that comprise it, such as, embryo, seed coat, and (or) pericarp. The sunflower dormancy can be reduced during after-ripening and by removing seed constraints. Our objective was to study how the conditions of storage and removal of the pericarp affect the level of dormancy in line A-3. Also we provide insight on the basis of the morphological characteristics of A-3 pericarp-imposed dormancy. A germination test was conducted on dry cypselas with and without pericarp, at 30 and 70 days after harvest. For histological analysis, permanent slides of pericarp cross-sections were obtained. The germination percentage showed significant differences between cypselas with intact pericarp (30 days after harvest = 26%; 70 days after harvest = 77%), and cypselas without pericarp (30 days after harvest = 65%; 70 days after harvest = 96%). This indicates that the pericarp plays an important role in regulating physical dormancy in the seed of sunflower line A-3, and that its relative contribution to the dormancy level is modified during after-ripening.

Early Response of Defense Related Genes to Secondary Downy Mildew Infection in Sunflower Line with Pl6 Gene

Sunflower line with resistance for downy mildew (Ha-26R) and susceptible line (Ha-26S) were inoculated with the suspension of

Influence of biostimulators on germination parameters and early growth of sunflower seedling

The aim of this paper was to examine the influence of biostimulators, with or without the use of fungicides, on seed quality parameters and early growth of sunflower seedling. Testing was conducted at the Institute of Field and Vegetable Crops in Novi Sad on cytoplasmic male sterile sunflower line HA-26-PR. The experiment included six treatments: Slavol S (25%), Slavol S (25%) + fungicide (a.m. metalaxyl-M), Bioplant Flora (2%), Bioplant Flora (2%) + fungicide (a.m. metalaxyl-M), Slavol S (25%) + Bioplant Flora (2%), Slavol S (25%) + Bioplant Flora (2%) + fungicide (a. m. metalaxyl-M) and control. Results of the research showed that treating the seed with biostimulators prior to sowing has a significant influence on seed germination parameters and early growth of sunflower seedling. The individual application of fertilizers, with or without the use of fungicide, gave a positive effect, while their combination led to a negative effect. The individual application of fertilizers, with or without combination with fungicide, may increase germination energy by 5%, germinability by 6%, but may also reduce the time of seed germination. Likewise, their use has a positive effect on early growth of sunflower seedling.