Tissue Culture Response from Seedling Explants of Commercial Barley Cultivars Grown in Bulgaria

Abstract

Only a few studies have been conducted in which regenerability of barley has been examined. In the current study, 17 barley genotypes (nine two-row malting type: Aster, Emon, Ruen, Jubiley, PV101, Körten, Krassi 2, Perun and Igri, and eight six-row feed types: Karnobat, Hemus, Jerun, Veslets, Aheloi 2, Diana, Panagon, and Izrgev) were evaluated for tissue-culture response from seedlings during a three-year period. Regenerable calli were obtained from all tested genotypes. Although there was much variation in regeneration capacity among the cultivars, plants were obtained from all cultivars. The majority of green plants grown to maturity were fertile and normal in appearance. The frequency of embryogénie callus induction and regenerability was influenced by genotype and growth conditions of the donor plants. Genotype was the most important determinant of the in vitro response. The best in vitro performance, based on ability to form regenerable calli and to regenerate plants was observed for Ruen, Aster, and Emon.     

The contribution of mammary infections by coagulase-negative staphylococci to the herd bulk milk somatic cell count

Quarter foremilk samples were taken at 2–3 weekly intervals for several years in a experimental herd comprising about 45 cows. The samples were submitted to bacteriological analysis and somatic cell counting. The most prevalent quarter infections from 1982 to 1988 were by coagulase-negative staphylococci (15–20% of all the quarters sampled). Most of these (75.6%) persisted until drying-off Dry cow therapy eliminated 86.5% of these infections. Comparison of udder quarters within cows, involving 775 samples from pairs of non-infected quarters and quarters infected by coagulase-negative staphylococci, yielded geometric means of somatic cell counts of 210 000 and 420 000 cells/ml, respectively. The correlation (r=0.87) between the herd bulk milk somatic cell count (SCC) and its estimation from the quarter milk somatic cell count performed on the same day allowed us to evaluate the contribution of the different categories of quarters, according to their infection status, to the herd bulk milk SCC. Quarters infected by a major pathogen (8.5% of samples) gave rise to 46.6% of the total number of cells, while quarters infected by coagulase-negative staphylococci (17.8% of samples) gave rise to 18.1%. Although coagulase-negative staphylococci represented only a secondary source of somatic cells as compared to major pathogens, they were not a negligible source considering the threshold of 300 000 somatic cells advocated for herd milk of good quality.     

不同管理条件下奶牛隐性乳房炎发生初探

乳汁中的体细胞增多，特别是白细胞增多是乳房炎的特点之一，因此，根据乳中体细胞数含量的多少来判断是否患有隐性乳房炎是一种可靠的检测手段本试验对不同管理状况下的奶牛随机抽样以检测乳汁中的体细胞数．结果显示：来自管理状况较好的合肥某乳业公司样本中的体细咆数明显低于安徽农业大学校牧场样本的体细胞数．两样本相比较，前者隐性乳房炎的发病率(30％)低于后者的发病率(50％)。     

Caprine somatic cell nuclear transfer using in vivo matured oocytes collected by laparoscopic follicular aspiration

In vivo matured oocytes collected by laparoscopic follicular aspiration (LFA) from hormone treated female goats were used as recipient ooplasts for somatic cell nuclear transfer (SCNT). Japanese native (Shiba) goats were used as donor females and some donor females were used repeatedly (two or three times) at intervals of a few months. To induce synchronization of estrus, a sponge containing 0.5 g of progesterone was inserted into the vagina of each goat for 14 days. These animals were also treated with follicle stimulating hormone (FSH) in a series of 8 injections over 4 days. The first FSH injection was administered on the morning of day 9 of sponge insertion. On the morning of day 13, 50 µg of gonadotropin‐releasing hormone (GnRH) was injected into each animal. Twenty‐nine hours after GnRH injection, LFA was performed. After removal of cumulus cells, collected oocytes with the first polar body were selected and enucleated for nuclear transfer. Anterior pituitary cells isolated from an adult male Shiba goat were transfected with a DNA fragment containing the enhanced green flourescent protein gene and the puromycin resistance gene. A single donor cell was inserted into the perivitelline space of each enucleated oocyte and fusion was induced with one electric pulse of 20 V for 10 µs. The SCNT goat eggs were cultured in chemically defined medium at 38.5°C in 5% CO₂, 5% O₂, 90% N₂ for 9 days. By LFA, 396 oocytes were collected from a total of 30 females. After removal of cumulus cells, 64% of them extruded the first polar body. The percentage of SCNT goat eggs produced using in vivo matured oocytes which developed to the blastocyst stage (20–21%) was significantly higher (P < 0.05) than that produced with in vitro matured oocytes (3–8%).     

落叶松体细胞胚胎发生研究进展

综述了欧洲落叶松、日本落叶松、欧日和日欧杂种落叶松、西部落叶松和华北落叶松体细胞胚胎发生的研究进展，对胚性培养物的超低温保存、原生质体培养和基因转化等技术进行了介绍，并对落叶松体细胞胚胎发生的研究趋势作了展望。     

Cryopreservation of yellow-poplar and sweetgum embryogenic cultures

Cryopreservation has become anessential tool for operational application offorest tree embryogenic cultures, due to thelong evaluation periods needed for treesregenerated from these cultures. Fiveyellow-poplar (Liriodendron tulipifera)and seven sweetgum (Liquidambar spp.)embryogenic culture lines werestored in liquid nitrogen for 48 hours, afterwhich they were thawed and tested for regrowthand ability to produce somatic seedlings.Combinations of two sorbitol pretreatments andthree dimethylsulfoxide (DMSO) cryoprotectantlevels were evaluated for their impact onrecovery following cryogenic storage. The bestresults were obtained with 0.4 M sorbitol and5% DMSO, which provided 100% recovery.Somatic seedlings were regenerated from allculture lines and treatments, except for atransgenic sweetgum line.     

蔗糖对水曲柳体细胞胚发生的影响

以水曲柳子叶为外植体,研究了不同浓度的蔗糖对水曲柳愈伤组织和体细胞胚发生以及体细胞胚数量、质量的影响。结果表明:当NAA多于BA时,附加7%蔗糖的体细胞胚发生率最高;当BA多于NAA时,附加10%蔗糖的体细胞胚发生率最高,但每个外植体上体细胞胚的数量和质量不是最好的。综合认为,选定9%~12%的蔗糖浓度最有利于水曲柳体细胞胚的诱导。     

Influence of race and crossbreeding on casein micelles size

Casein (CN) micelles are colloidal aggregates of protein dispersed in milk, the importance of which in the dairy industry is related to functionality and yield in dairy products. The objective of this work was to investigate the correlation of milk CN micelles diameter from Holstein and Zebu crossbreds with milk composition (protein, fat, lactose, total and nonfat solids and milk urea nitrogen), somatic cell count (SCC), age, lactation stage and production. Average casein micelles diameters of milk samples obtained from 200 cows were measured using photon correlation spectroscopy and multiple regression analysis was used to find relationship between variables. CN micelle diameter, SCC and nonfat solids were different between animals with different Holstein crossbreed ratios, which suggests influence of genetic factors, mammary gland health and milk composition. Overall, results indicate the potential use of CN micelle diameter as a tool to select animals to produce milk more suitable to cheese production.     

The use of COLD‐PCR,DHPLC and GeneScanning for the highly sensitive detection of c‐KIT somatic mutations in canine mast cell tumours

The conventional polymerase chain reaction (PCR)/sequencing methods may be poorly suited for the detection of somatic mutations in canine mast cell tumour (MCT) samples owing to limited sensitivity. This study was aimed at establishing novel and more sensitive methods, assessing their limit of detection and comparing their sensitivity with conventional methods.Two different ‘driver’ somatic mutations of c‐KIT, together with the wild‐type counterparts, were cloned in plasmids to prepare standard samples with known concentrations of mutated alleles in a background of wild‐type alleles; the plasmids standards were assayed using either conventional or novel, highly sensitive technique. Conventional PCR/sequencing showed a sensitivity of 50–20%. Conversely, all the novel methods obtained higher sensitivities allowed reaching as low as 2.5–1.2% of the mutated DNA.The study demonstrates that early conventional methods could likely have underestimated the prevalence of KIT mutations of MCTs, therefore affecting the assessment of their relevance in prognosis and tyrosine kinase inhibitor (TKI) treatment effectiveness.