培养三倍体柑桔植株的研究

以柑桔异源四倍体体细胞杂种（哈姆林甜橙（ＣｉｔｒｕｓｓｉｎｅｎｓｉｓＯｓｂｅｃｋｃｖ．Ｈａｍｌｉｎｓｗｅｅｔｏｒａｎｇｅ）＋粗柠檬（Ｃ．ｊａｍｂｈｉｒｉＬｕｓｈｃｖ．Ｒｏｕｇｈｌｅｍｏｎ）为父本与单胚的二倍体柑桔类型（宜昌橙（Ｃ．ｉｃｈａｎｇｅｎｓｉｓＳｗｉｎｇｌｅ）与华农本地早（Ｃ．ｒｅｔｉｃｕｌａｔａＢｌａｎｃｏｃｖ．Ｈｕａｎｏｎｇｂｅｎｄｉａｚｏ）的杂交后代）杂交。通过幼胚早期离体培养,获得     

哈姆林×路比血橙幼胚组培植株高接结果观察

哈姆林×路比血橙幼胚组培植株高接结果观察赵琦，韦党扬，李兴忠，李向林，赵国柄（贵州省柑桔研究所罗甸５５０１００）在柑桔杂交育种工作中，由于受种子多胚的干扰，杂交后很难区分和获得杂种苗．早期离体培养幼合子胚，可使其脱离珠心胚的干扰而单独发育成苗，提高杂...     

试管嫁接提高柑桔三倍体组培苗成苗率试验

选育三倍体类型是柑桔育种的热点之一。四倍体与二倍体杂交是获得三倍体的一条有效途径。但三倍体胚有早期败育的现象,为了获得三倍体植株,幼胚须进行早期离体培养。本实验室以异源四倍体柑桔体细胞杂种为父本与二倍体有籽品种或类型杂交,通过幼胚早期离体培养,获得了一批三倍体植株。但是,由于三倍体幼胚离体培养再生的试管苗大部分生长势很弱,生长慢,根发育不良,难以形成完整植株,即使成株,移栽成活率也较低。为了提高三倍体幼胚成苗率及移栽成活率,获得生活力强的三倍体材料,我们进行了试管微嫁接及不同时期移栽的试验。１材…     

三倍体柑桔幼胚离体培养研究

三倍体柑桔幼胚离体培养研究伊华林１邓秀新２史永忠２郭文武２（１华中农业大学园艺系；２华中农业大学作物遗传改良国家重点实验室，武汉４３００７０）关键词柑桔；三倍体；体细胞杂种；胚抢救；有性杂交ＳｔｕｄｉｅｓｏｎＣｕｌｔｕｒｅｏｆＩｍｍａｔｕｒｅＴｒｉ...     

我国柑桔离体培养研究的进展

近20年来柑桔离体培养研究有了飞跃发展,本文试就国内在这方面的研究作一总结。 一、胚胎培养 柑桔杂交育种最大的障碍是种子的多胚性,早期离体培养幼合子胚,可脱离珠心胚的干扰而单独发育,以提高杂种率。国内外对柑桔合子胚的早期离体培养研究,至今尚未取得令人满意的结果。陈振光等用雪柑×(椪柑+福桔)     

特早熟杏胚培养试验

果树极早熟、早熟品种胚发育不全 ,其种子采用常规播种育苗的发芽率极低 ,而采用胚培养技术 ,不仅能提高胚的萌发率、成苗率 ,而且可望选育出成熟期更早的新品种。目前 ,桃、樱桃、李、梅等核果类果树多个早熟品种胚培养已经成功 [1～ 4 ] ,杏早熟品种幼胚培养国内外报道较少 [5～ 7] ,本试验以杏极早熟品种红荷包幼胚和胚珠为试材 ,进行了培养试验 ,现将结果报道如下。1　材料与方法　　供试材料为杏特早熟品种红荷包自然杂交果实的胚珠、幼胚 ,试材摘自山西省农业科学院园艺研究所杏品种资源圃 ,于盛花后 45天(半硬核期 )采样。胚珠培养　…     

离体葡萄未成熟胚成苗途径研究

以巨峰葡萄剥离幼胚及红井川品种切喙胚珠为外值体接种至含不同浓度２,４Ｄ、ＮＡＡ与６－ＢＡ配比的ＮＮ－１９６９培养基中,诱导葡萄未成熟胚产生胚状体及实现胚萌发。两品种均是在含２,４－Ｄ０．５ｍｇ·１－１、６－ＢＡ１．０ｍｇ·１－１的培养基中诱导胚进入胚状体发生途径的;而在含ＮＡＡ（１．０、２．０ ｍｇ·１－１）与 ６－ＢＡ（０．２、１．０）或２．０ ｍｇ·１－１ＺＴ 配比的培养基上幼胚进入胚萌发途径成苗。由此讨论了培养基激素、胚发育时期对胚培养两种途径的诱导和控制。     

山楂胚离体培养及四倍体诱导研究

以山楂为材料,研究了幼胚和成熟胚离体培养及四倍体诱导技术。花后40 d的山楂幼胚离体培养的成苗率很低,供试的15份资源的成苗率都小于20%。成熟胚离体培养的成苗率较高,调查的4份资源的成苗率均大于50%。用0.5%秋水仙碱+1%二甲基亚砜的混合溶液处理山楂成熟胚48 h,然后在SC+IBA 0.1 mg.L-1+TDZ 1.0 mg.L-1培养基上诱导芽分化。对秋水仙碱处理成熟胚再生植株进行茎尖染色体数目观察,检测的67个隆化粉肉植株中有3个是四倍体,变异率达4.5%。研究建立了山楂多倍体离体诱导体系,为培育山楂多倍体品种奠定了重要基础。     

荔枝叶片愈伤组织的诱导

已报道的荔枝离体培养主要取材是幼胚或花药，对叶片的离体培养未见报道。本试验对荔枝叶片的培养作了一些探索。     

黑核桃体细胞胚状体发生及其基因转化系统

以我国种植的优良品种黑核桃（Ｊｕｇｌａｎｓ ｎｉｇｉａ Ｌ．）幼胚和幼叶为外植体诱导出体细胞胚状体，通过根癌农杆菌介导将ｎｐｔⅡ和ｇｕｓ基因导入细胞胚，同时研究了激素、体细胞胚的发育时期及预培养等对转化率的影响，建立了黑核桃体细胞胚基因转化系统，还分析了体细胞胚胎转化系统的特点和潜力。     

柑橘体细胞杂种有性后代花粉母细胞减数分裂的观察

柑橘体细胞杂种后代为三亲来源,采用酸解压片法对柑橘体细胞杂种[哈姆林甜橙(Citrus sinensis Osbeck)+粗柠檬(C.jambhiri Luss)]与单胚二倍体植株宜本杂4号[华农本地早橘(C.reticulata Blanco)×宜昌橙(C.ichangensis Swingle)]的有性杂种后代进行减数分裂观察,以明晰柑橘体细胞杂种有性后代减数分裂行为。结果表明,有性后代二倍体后代减数分裂行为与二亲杂种后代相似;三倍体后代减数分裂过程异常现象较二亲杂种后代复杂,中期Ⅰ出现56.9%的不配对染色体以及各种多价体,中期Ⅰ和中期Ⅱ观察到游离在赤道板外的染色单体,变幅为1～3个,平均比例分别为51.2%和38.68%;后期Ⅰ和后期Ⅱ观察到染色体桥及染色体断片、落后染色体;四分体时期含微核等。同时对后代植株花蕾直径与花粉母细胞减数分裂各个时期之间的关系进行了分析。     

Shoot zinc (Zn) concentration varies widely within Brassica oleracea L. and is affected by soil Zn and phosphorus (P) levels

Summary

The low availability of zinc (Zn) in soils and crops affects dietary Zn intake worldwide. This study sought to determine if the natural genetic variation in shoot Zn concentrations ([Zn]shoot) is sufficient to pursue a crop improvement breeding strategy in a leafy vegetable crop. The gene-pool of Brassica oleracea L. was sampled using a large (n = 376) diversity foundation set (DFS), representing almost all species-wide common allelic variation, and 74 commercial varieties (mostly F₁). The DFS genotypes were grown at low and high soil phosphorus (P) levels under glasshouse and field conditions, and also in a Zn-deficient soil, with or without Zn-fertilisation, in a glasshouse. Despite the large variation in [Zn]shoot among genotypes, environment had a profound effect on [Zn]shoot. The heritability of [Zn]shoot was significant, but relatively low, among 90 doubled-haploid (DH) lines from a mapping population. While several quantitative trait loci (QTL) associated with [Zn]shoot occurred on chromosomes C2, C3, C5, C7, and C9, these were generally weak and conditional upon growth conditions. Breeding for [Zn]shoot in B. oleracea is therefore likely to be challenging. Shoot P concentrations increased substantially in all genotypes under low soil Zn conditions. Conversely, only some genotypes had increased [Zn]shoot at low soil P levels. Sufficient natural genetic variation may therefore exist to study some of the interactions between Zn and P nutrition.     

几种砧木对哈姆林甜橙植株生长、产量及果实品质的影响

以7种不同砧木的8年生哈姆林甜橙为试材,对植株营养生长、产量和果实品质进行了比较。结果表明,不同砧木对哈姆林甜橙生长和产量有不同影响,其中以卡里佐枳橙为砧的植株树冠体积最大,以光皮酸橘、枸头橙和李齐16-6枳为砧的植株树冠体积较小,单位树冠体积产量以光皮酸橘为砧的植株最高,单株挂果数则以李齐16-6枳和卡里佐枳橙为砧的最多。不同砧木对果实品质有显著影响,其中以光皮酸橘为砧的果实体积和单果质量最大,以兰普来檬为砧的果汁含量最高,以李齐16-6枳和卡里佐枳橙为砧的果实可溶性固形物(TSS)含量最高,以李齐16-6枳为砧的果实可滴定酸含量亦最高,而李齐16-6枳和兰普来檬为砧的果实维生素C含量最高。从外观品质看,枸头橙为砧的果实油斑病发生程度最高。综合评价认为,以卡里佐枳橙、李齐16-6枳为砧的单株具产量高、果汁TSS含量高和风味浓郁等优点,是哈姆林甜橙较为理想的砧木。     

Astragalus polysaccharides protects against free fatty acid-induced human vascular endothelial cell dysfunction via AMPK-eNOS pathway

AIM: To study the protective effect of Astragalus polysaccharides (APS) on free fatty acid-induced injury in human umbilical vein endothelial cells (HUVECs). METHODS: Cultured HUVECs were divided into control group, APS group [APS (200 mg/L) treated for 24 h], free fatty acid group [free fatty acid (0.25 mmol/L) treated for 24 h], free fatty acid plus APS group [free fatty acid (0.25 mmol/L) and APS (200 mg/L) treated for 24 h], and compound C group [free fatty acid (0.25 mmol/L) and APS (200 mg/L) and AMPK inhibitor compound C (10 μmol/L) treated for 24 h]. The cell viability was detected by MTT assay. Nitric oxide (NO) content in the medium was determined by nitrate reductase assay. The protein levels of total adenosine monophosphate-activated protein kinase (AMPK), phosphorylated adenosine monophosphate-activated protein kinase (p-AMPK), endothelial nitric oxide synthase (eNOS) and phosphorylated endothelial nitric oxide synthase (p-eNOS) were measured by Western blot. RESULTS: No significant difference of all indexes between APS group and control group was observed. The cell viability in free fatty acid group decreased significantly compared with control group. The cell viability in free fatty acid plus APS group was significantly improved as compared with free fatty acid group. The cell viability in compound C group was almost the same as that in free fatty acid group. The No content and protein levels of p-AMPK and p-eNOS in free fatty acid group decreased obviously as compared with control group, while the NO content and protein levels of p-AMPK and p-eNOS in free fatty acid plus APS group increased obviously compared with free fatty acid group. No significant difference of the p-AMPK and p-eNOS protein levels between free fatty acid plus APS group and free fatty acid group was observed. No significant difference of the AMPK and eNOS protein levels in all groups was found. CONCLUSION: APS attenuates the free fatty acid-induced injury, and its mechanism is related to the AMPK-eNOS signal pathway.     

Combination of R848 and Poly(I: C) promotes antigen presentation by DC and enhances killing effect of CTL on lung adenocarcinoma A549 cells

AIM: To investigate the effect of R848 (a Toll-like receptor 7/8 agonist) combined with poly-inosinic:polycytidylic acid[Poly(I:C), a Toll-like receptor 3 agonist] on dendritic cell (DC) maturation, and the killing effect of DC-induced cytotoxic T-lymphocytes (CTL) on human lung adenocarcinoma A549 cells. METHODS: Mononuclear cells were isolated from human peripheral blood and induced to differentiate into DC. The whole-cell lysate of A549 cells, namely tumor cell lysate (TCL), was used as antigen. R848 combined with Poly(I:C) was used as adjuvant to stimulate the DC. DC surface markers were analyzed by flow cytometry. The DC stimulated by antigen was co-cultured with T-lymphocytes for 7 d to induce CTL. The culture supernatant and CTL were collected. The levels of interleukin-12 (IL-12) p70, interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) in the supernatant were measured by ELISA. The CTL and A549 cells were co-cultured for 16 h, and the cytotoxicity was observed by LDH assay.RESULTS: The expression of CD83 and CD80 on the DC surface, and the secretion of IL-12 p70 in DC-R848+Poly(I:C) group were significantly increased compared with DC-TCL group (P<0.01). In addition, the cytotoxicity of CTL for A549 cells in DC-R848+Poly(I:C) group was significantly enhanced compared with DC-TCL group (P<0.01). The secretion levels of IFN-γ and TNF-α in DC-R848+Poly(I:C) group were significantly elevated compared with DC-TCL group (P<0.01). CONCLUSION: R848 combined with Poly(I:C) significantly promotes DC maturation and activation, and enhances the antigen-presenting effect of DC and the cytotoxicity of DC-induced CTL.     

柑橘及其近缘属植物LFY同源基因的比较

LFY基因处于成花调控网络的关键位置,不仅调控开花时间和花转变,而且在花序和花的发育中也起重要作用。为了进一步探讨柑橘及其近缘属植物开花的分子机理,利用PCR技术分别从兴津温州蜜柑(Citrus unshiuMarcovitch)、无核椪柑(Citrus reticulata Blanco)、沙田柚[Citrus grandis(L.)Osbeck]、融安金柑(Fortunella crassifoliaSwing)和无核黄皮[Clausena lansium(Lour.)Skeels]叶片中分离克隆了LFY全长同源基因。结果表明兴津温州蜜柑、无核椪柑、沙田柚、融安金柑和无核黄皮中的LFY全长同源基因的核苷酸长度分别为2090、2086、2092、2081、2089bp,分别编码398、398、398、398和397个氨基酸,这些同源基因均由3个外显子和2个内含子组成。同源性分析发现,这些LFY全长同源基因的核苷酸序列和氨基酸序列同源性高,分别为92%～99%和95%～100%。亲缘关系分析结果与当前的植物学分类结果一致。     

Erythropoietin inhibits eryptosis induced by reactive oxygen species

AIM: To observe the influence of erythropoietin (EPO) on eryptosis and production of reactive oxygen species (ROS) in erythrocytes under stimulation of hydrogen peroxide (H₂O₂),and to explore its related mechanism. METHODS: The erythrocyte suspension (1%) was cultured in vitro and divided into 3 groups:control group (C group, the culture medium was PBS), H₂O₂ group (H group, the culture medium was PBS containing H₂O₂ at final concentration of 100 μmol/L) and EPO group (E group, the culture medium was PBS containing H₂O₂ at final concentration of 100 μmol/L and EPO at final concentration of 2×10⁴ U/L). The erythrocytes were collected at 24 h and 60 h. The eryptosis was detected by flow cytometry with Annexin V staining. The production of ROS and intracellular calcium ion concentration ([Ca²⁺]_i) were also analyzed by flow cytometry. RESULTS: The eryptosis in C group was increased as the incubating time extended. The eryptosis in H group was higher than that in C group (P<0.01), while that in E group was lower than that in H group (P<0.01). Meanwhile, ROS production and[Ca²⁺]_i were higher in H group than those in C group (P<0.01), but those were lower in E group than those in H group (P<0.05 or P<0.01). CONCLUSION: EPO inhibits eryptosis induced by H₂O₂ and its mechanism may be related to antioxidant effect and change of[Ca²⁺]_i.     

Investigation of amino acid sequence of specific ouabain conjugated peptides

AIM: The purpose of the study was to investigate the gene and amino acid sequence of specific ouabain conjugated peptides (OCP) in order to get an experimental bases to block or antagonist the actions between endogenous ouabain(EO) and sodium pump in hypertension. METHODS: Screening the phage displayed 12-peptide library by biopaning for OCP. The sequence of each selected peptide was determined and the sequence was analyzed through internet. The bioactivity was determined by erythrocyte [⁸⁶Rb] uptaking. RESULTS: Three kinds of peptides were screened out. Peptide A (12 peptide) was occupied in 66.7%(8/12), peptide B (8 peptide) 16.7% (2/12) and peptide C (12 peptide) 8.3% (1/12). There was only one case without insertron. The analysis of protein showed that there were no homogenous between peptide A, B, C and sodium pump. The amino acid sequence of specific OCP was Leu-Leu-Ala-Asp-Thr-Thr-His-His-Arg -Pro-Trp-Thr. CONCLUSIONS: Determination of the sequence of OCP supplies an important experimental foundation for ouabain research. The results also show that phage display peptide library is an effective, simple and efficient method to select specific steroid receptors.     

辐射花粉授粉诱导甜瓜单倍体

为了掌握辐射花粉授粉诱导甜瓜单倍体技术,以5种基因型甜瓜为材料,通过应用γ射线辐射的花粉进行授粉结合胚胎培养,从2种基因型中获得了单倍体植株,单倍体的平均诱导频率为0.29%。这2种基因型分别属于厚皮和薄皮甜瓜类型。花粉的辐射剂量为300Gy和600Gy时,供试材料的平均坐果率分别为50%和10%。辐射剂量为300Gy时,诱导厚皮甜瓜4810和薄皮甜瓜新富玉形成了单倍体植株,单倍体植株的诱导频率分别为0.55%和0.63%;剂量为600Gy时未能诱导供试材料形成单倍体。研究结果表明母本的基因型和花粉的辐射剂量对坐果率和单倍体的产生有一定的影响。此外,对单倍体植株与正常二倍体植株的形态学进行了比较观察,结果表明单倍体植株生长势弱,叶片较小,雄花和雌花均败育。     

秦岭地区春兰和蕙兰的花挥发性成分研究

采用顶空固相微萃取和气相色谱-质谱联用技术（HS–SPME–GC–MS）对原产于秦岭不同地点的春兰和蕙兰的鲜花进行了挥发性成分测定。结果表明该地区春兰的花中挥发性成分有40多种,主要成分有3–乙基–2–甲基–1,3–己二烯、(E)–2–辛烯醛和2–壬烯醛等;不同产地的春兰花中主要的挥发性物质构成比较类似,花香类型比较单一,且多数为无香型。而该地区蕙兰的花中挥发性成分多达50种以上,主要成分有(E)–橙花叔醇,二十二碳六烯酸和[1à,2à(Z)]–茉莉酸甲酯等。蕙兰花中挥发性成分中大多为有芳香味的物质,从而使得蕙兰的花具有更浓郁的香味。此外,蕙兰花挥发性成分构成也比春兰复杂,因此比春兰具有更多的花香类型。