凤尾蕨科植物蜈蚣草的组织培养

[目的]对凤尾蕨科植物蜈蚣草的组织培养方法进行研究。[方法]以野外采集的叶芽为外植体,以1/2 MS＋3%蔗糖＋0.7%琼脂为基本培养基,通过激素配比试验,筛选蜈蚣草愈伤组织诱导和继代培养的培养基配方。[结果]从外植体诱导出愈伤组织最佳培养基配方为1/2 MS＋3%蔗糖＋0.7%琼脂＋0.5 g/L PVP＋0.1 mg/L KT＋0.5 mg/L 2,4-D;从愈伤组织诱导出GGB和从GGB诱导出幼苗最佳培养基配方为1/2MS＋3%蔗糖＋0.7%琼脂＋0.5 g/L PVP＋1.0 mg/L KT＋0.01 mg/L 2,4-D;另外,使用1/2 MS＋3%蔗糖＋0.7%琼脂＋0.5 g/L PVP＋0.5 mg/L 2,4-D做继代培养可以使愈伤组织持续增殖。[结论]研究直接使用野外材料组织培养,简化了试验步骤,是一种方便快速的蜈蚣草组培方法。     

Molecular cloning and expression of kidney‐type glutaminase from common carp (Cyprinus carpio) and its up‐regulation by glutamine in primary culture enterocyte

Glutaminase (GLS) is the key enzyme of glutamine (Gln) metabolism and utilization. In this study, a cDNA encoding GLS protein was identified from common carp Cyprinus carpio intestine. The open reading frame of GLS cDNA encodes a polypeptide of 595 amino acids, which shows a high similarity with its zebrafish Danio rerio counterpart. Bioinformatic analysis showed the protein belongs to kidney‐type GLS. The putative protein has glutaminase domain and ankyrin repeats domain, which are highly conserved among vertebrate orthologues. Real‐time quantitative PCR analysis revealed that the abundance of GLS mRNA was the highest in the white muscle, followed by the brain, eyeball and pituitary. Glutaminase was ubiquitously expressed in all intestinal segments of common carp. The activity of GLS did not distribute uniformly along the entire length of the intestine. In primary culture enterocyte, and the expression of GLS mRNA is up‐regulated quickly and effectively by Gln.     

Characterization of tuberculous granulomas in different stages of progression and associated tertiary lymphoid tissue in goats experimentally infected with Mycobacterium avium subsp. hominissuis

Oral infection of goats with Mycobacterium avium subsp. hominissuis (MAH) resulted in a large variety of granulomas in organized gut-associated lymphatic tissues and intestinal lymph nodes. To characterize the cellular composition of granulomas, CD4⁺, CD8⁺, γδ, B lymphocytes and plasma, CD25⁺, CD68⁺, MHC-II⁺, Ki67⁺ and endothelial cells were labeled in consecutive frozen sections by immunohistochemistry and acid fast bacilli (AFB) by Kinyoun stain. Granulomas with extensive necrosis, little mineralization and variable numbers of AFB surrounded by many CD4⁺ T cells, but only few epitheloid macrophages were observed in severely sick goats at 2–3 mpi. They were interpreted as exuberant immune reaction. Organized granulomas with very few AFB were seen in clinically healthy goats at 13 mpi. The necrotic cores were surrounded by a zone of granulomatous infiltrate with many epitheloid macrophages and few lymphocytes. This zone was initially wide and highly vascularized and became progressively smaller. It was enclosed by an increasing layer of connective tissue. All organized granulomas were surrounded by compartimentalized tertiary lymphoid tissue. The granulomas in experimental infection of goats with MAH reflect the heterogeneity of lesions seen in mycobacterial infections of humans and ruminants and are therefore valuable for comparative research.     

Physiological and pathological roles of mast cells in adipose tissue

Mast cells are important cells for the innate immunity that reside in tissues including adipose tissue and are involved in various physiological and pathological processes by producing a range of biological mediators. Adipose tissue not only acts as an energy depot and regulator of energy homeostasis that can deposit excess energy and dissipate energy through heat, but also is an active endocrine organ capable of producing hormones and adipokines. The dysfunction of adipose tissue is highly correlated with metabolic disorders, such as obesity, type 2 diabetes mellitus and so on. This review focuses on the physiological and pathological roles of mast cells in adipose tissue.     

Advance in study of batokines

Brown adipose tissue is a type of adipose tissue and is present in all mammals. It is not only the main site for adaptive thermogenesis in vivo, but also secretes and releases many cytokines in the form of endocrine to regulate a variety of metabolic processes and prevent and treat obesity and metabolic diseases. Therefore, identification of the types and effects of brown adipocytokines (batokines) may be crucial for the treatment of a variety of obesity-related metabolic diseases. Based on the latest research progress, batokines and their functions are reviewed.     

利用绿色荧光蛋白优化辣椒遗传转化体系

【目的】农杆菌介导的辣椒遗传转化较为困难,至今仍未建立高效转化体系。绿色荧光蛋白(GFP)基因是植物遗传转化中常用的报告基因,以GFP为报告基因,优化农杆菌介导的辣椒遗传转化体系。【方法】利用GFP表达系统,统计3个辣椒品种(‘HP’‘8214’和‘L55’)中3种不同外植体(子叶、下胚轴和Flamingo-bill外植体)的不定芽分化率、不定根分化率与荧光阳性率,探究农杆菌侵染浓度、侵染时间、预培养时间和共培养时间等因素对不定芽、不定根分化率及荧光阳性率的影响。【结果】3个辣椒品种的Flamingo-bill外植体不定芽分化率均显著高于下胚轴与子叶外植体,其中‘L55’ Flamingo-bill外植体的不定芽分化率最高、达77.59%,故选用‘L55’ Flamingo-bill外植体进行后续研究。4种农杆菌不同侵染浓度和时间组合下,‘L55’Flamingo-bill外植体均可产生不定芽、不定根及表达GFP的愈伤组织,当农杆菌侵染浓度为OD₆₀₀=0.05,侵染时间为30 min时,不定芽分化率和荧光阳性率最高,分别达48.39%、4.84%。在6种不同预培...     

咖啡小粒种六倍体和中小粒杂种四倍体胚乳的组织培养

进行了利用咖啡胚乳离体培养技术培育六倍体咖啡胚乳植株及四倍体中小粒种咖啡种间杂种的研究。结果表明:将小粒种咖啡自花授粉所得六倍性胚乳及以中粒种咖啡为母本、小粒种咖啡为父本进行杂交所获得的四倍性种间杂种胚乳,分别在附加有6-BA1~2mg/L和NAA2mg/L的改良MS培养基上培养,能诱导出有效的愈伤组织;在附加有6-BA或KT0.5mg/L,NAA0.1mg/L的改良MS培养基上,愈伤组织能分化出各种类型的胚状体;在附加有NAA或IBA0.5mg/L及2g/L活性炭的1/2MS培养基上,胚状体可被诱导生根而再生完整的六倍体咖啡胚乳植株和四倍体种间杂种咖啡胚乳植株。      

菊叶薯蓣组织培养及快速繁殖

菊叶薯蓣组织培养以大田栽培苗地上部分幼嫩茎段作外植体。芽诱导培养基为MS+6-BA0.5～2.0mg·L-1+NAA0.1mg·L-1,继代增殖培养基为MS+6-BA1.0mg·L-1+NAA0.1 mg·L-1,生根培养基1/2MS+NAA0.5mg·L-1,每个过程均培养30d,每节段增殖2～3倍,生根率为85%以上。移栽基质为土壤∶珍珠岩(1∶3),遮光率为80%,保持较高湿度,每7d喷施1/2MS无机盐,1个月后成活率可达60%～80%。     

Micropropagation of Prunus scoparia,a Suitable Rootstock for Almond under Drought Conditions

Prunus scoparia is a wild deciduous shrub, usually living on dry calcareous soils of the rocky mountains and has been used as a grafting rootstock for domesticated almonds to provide drought resistance. In the current study, micropropagation ability of P. scoparia was investigated using cytokinin and auxin. Uniform nodal shoot pieces (3–5 cm in length) of seedlings were used as explants. The explants were disinfected with 10% sodium hypochlorite solution. For adventitious shoot induction and proliferation, Murashige and Skoog (MS) media containing 7.00 g/l agar and 30.00 g/l sucrose containing five concentrations of benzyl adenine (BA) (0.00, 0.50, 1.00, 2.00, and 4.00 mg/1) and also containing six concentrations of Thidiazuron (TDZ) (0.00, 0.50, 1.00, 2.00, 5.00, and 7.00 mg/1) were compared. For rooting, in vitro shoots (2–3 cm) were transferred into ½ MS medium supplemented with 30 g/l sucrose, 7.50 g/l agar, and different concentrations of IBA (0.00, 0.25, 0.50, and 1.00 mg/l) and NAA (0.00, 0.25, 0.50, and 1.00 mg/l). Based on the results obtained for shoot proliferation, only 2.00 and 4.00 mg/l BA and 2.00 mg/l TDZ concentrations generated shoots, while other treatments did not show shoot proliferation. Among the three treatments that generated shoots, the best results for shoot number, leaf number, and leaf color quality were observed in media containing 2.00 mg/l TDZ. Based on the results obtained for rooting, the effect of IBA concentrations on the rooting percentage, root number, and root length was significant. Among IBA concentrations, only 0.50 mg/l IBA induced rooting, while there was no rooting in the media containing other IBA concentrations. None of the NAA concentrations showed rooting. In conclusion, MS culture medium supplemented with 2.00 mg/l TDZ and ½ MS culture medium supplemented with 0.50 mg/l IBA are suggested for in vitro shoot proliferation and rooting of P. scoparia, respectively. The results presented herein could be used for in vitro selection and micropropagation of P. scoparia.