转rolA、B、C基因枳橙快繁技术

通过对转rol基因枳橙B、D、E系及对照的高接植株春梢茎段进行萌芽诱导、增殖及试管苗生根成苗的试验,摸索出适宜枳橙快繁各种培养基配方及培养条件。结果表明,以MS+6-BA1mg/L的培养基适合于芽萌发,以MS+6-BA0.5mg/L+NAA0.05mg/L的培养基适合于芽增殖,生根则以1/2MS+NAA0.5mg/L+0.2%活性炭的组合最佳,转基因各系生根率均在93.3%以上,显著高于对照生根率(66.7%)。快繁苗移栽成活率可达90%,目前生长良好。     

不同氮素形态对枳橙幼苗生长特性的影响

采用水培方法研究了不同形态氮素配比对枳橙[Citrus.sinensis(L.)Osb×Poncirus trifoliate(L.)Raf]幼苗生长特性的影响。结果表明,不同氮素形态配比营养液对枳橙幼苗的生长均有一定的促进作用,其中混合态氮素对植株地上部形态特征的影响好于单一态氮素,NO3-∶NH4+=5∶5处理更利于株高、茎粗和叶片数的增加;其次是NO_3~-∶NH_4~+=7∶3处理;单一态氮素形态处理中,全硝培养好于全铵培养。NO_3~-∶NH_4~+=5∶5处理促进了地下部主根的伸长和侧根的增加,却对主根粗度无明显影响。全铵培养对幼苗地上部和地下部生长均有抑制作用,甚至是毒害作用。不同氮素形态配比对枳橙幼苗叶绿素a、叶绿素b及叶绿素总量的影响趋势基本一致,混合态氮素对叶绿素a、叶绿素b和叶绿素总量的促进好于单一态氮素,且当处理为NO_3~-∶NH_4~+=5∶5时叶绿素含量持续增加,并且更利于其积累;而全铵处理则不利于叶绿素的增加与积累。     

Host range and symptomatology of a graft-transmissible pathogen causing bud union crease of citrus on trifoliate rootstocks

A graft-transmissible pathogen causing bud union crease of Nagami kumquat SRA–153 on Troyer citrange was characterized for host range and symptomatology. Buds of Marsh grapefruit, Nules clementine, Eureka lemon and Pineapple sweet orange preinoculated with kumquat SRA–153 were propagated on citrange rootstocks. Some plants of Nules clementine and Eureka lemon had developed bud union crease six months after propagation, whereas all Marsh grapefruit and Pineapple sweet orange plants still showed normal bud union after one year. On indexing these preinoculated species, Nules clementine and Eureka lemon caused vein clearing in Pineapple sweet orange and Dweet tangor, chlorotic blotching in Dweet tangor and stem pitting in Etrog citron, whereas Marsh grapefruit and Pineapple sweet orange caused only chlorotic blotching in Dweet tangor and stem pitting in Etrog citron. Following shoot-tip grafting in vitro of kumquat SRA–153, kumquats 38–1 and 497–2 obtained from it caused chlorotic blotching in Dweet tangor and stem pitting in Etrog citron, but not vein clearing in Pineapple sweet orange and Dweet tangor or bud union crease when propagated on citrange. These results suggest the presence of at least two pathogens or pathogen strains in kumquat SRA–153 and the elimination of one of them after shoot-tip grafting in vitro or inoculation on Marsh grapefruit or Pineapple sweet orange. They also indicate that the pathogens in kumquat SRA–153 can be detected by indexing on Dweet tangor or Etrog citron.     

卡里佐枳橙幼苗病害致病菌的分离和鉴定

在卡里佐枳橙幼苗上发生一种严重病害,为害卡里佐枳橙茎干,造成褐变,叶片干枯或脱落,严重时植株死亡。从卡里佐枳橙病害苗上分离纯化得到4株真菌,分别为MP1、MP2、MP3和MP4。致病性测定发现,上述4种真菌的致病程度不同,MP2孢子悬浮液处理发病最轻,MP3孢子悬浮液处理发病最为严重;对这4种菌进行形态学和分子生物学鉴定,发现3株属于链格孢属(Alternariasp.),另外1株属于盾壳霉属(ConiothyriumCorda)。     

枳、枳橙和甜橙实生苗的钾利用效率比较

对枳、枳橙和甜橙的钾利用效率进行比较,将枳、枳橙和甜橙实生苗置于不同钾浓度条件下生长,检测不同时间植株的生物量和钾含量。结果显示:无论在高钾(20 mmol/L K+)还是低钾(100μmol/L K+)条件下,枳橙根部干重都显著高于枳,枳橙根部钾含量亦显著高于枳。在高钾条件下,枳橙和冰糖橙根部干重无显著性差异,但枳橙根部钾含量显著高于冰糖橙;无论在高钾还是低钾条件下,枳、枳橙和冰糖橙冠部干重均没有显著性差异,冠部钾含量表现为冰糖橙枳橙枳。从植株钾利用效率而言,枳橙作为砧木应优于枳;枳橙作为枳与甜橙的杂种,在根部钾利用效率上亦高于甜橙。     

The effects of low boron on growth,gas exchange,boron concentration and distribution of ‘Newhall’ navel orange (Citrus sinensis Osb.) plants grafted on two rootstocks

The effects of low boron (B) on plant growth, photosynthesis, B concentration and distribution of ‘Newhall’ orange (Citrus sinensis Osb.) plants grafted on either Trifoliate orange (Poncirus trifoliata (L.) Raf.) or Carrizo citrange [C. sinensis (L.) Osb. × Poncirus trifoliata (L.) Raf.] rootstocks were investigated. One-year-old plants of the two scion-rootstock combinations were grown for 183 days in sand:perlite (1:1, v/v) medium under greenhouse conditions. The plants were irrigated with half-strength Hoagland's nutrient solutions containing four B concentrations (0.01, 0.05, 0.10 and 0.25 mg l⁻¹). The growth of root, stem of scion and leaves was less affected by low B treatments when ‘Newhall’ scion was grafted on Carrizo citrange than on Trifoliate orange. Thus, the growth of scions under low B conditions was mainly depended on the rootstock used, i.e., Carrizo citrange-grafted plants were more tolerant to low B compared to the plants grafted on Trifoliate orange. Boron concentrations in all plant parts decreased significantly by decreasing the B supply in the nutrient solution. Leaves were the dominant sites of B accumulation and showed the greatest reduction in B concentration compared to the other plant parts, as B concentration in the nutrient solution decreased. Irrespective of the rootstock, B levels in the upper–younger leaves were substantially higher than in basal-older leaves when plants were exposed to low B concentrations (≤0.05 mg l⁻¹), suggesting that under such conditions B was preferentially translocated to upper–younger leaves to support their growth. Furthermore, B distribution in different plant parts indicated that there was a restriction in translocation of B from root to scion tissues (stems and leaves of scion) under conditions of limited B availability. In addition, low B treatments decreased leaf photosynthetic rate, stomatal conductance and transpiration rate but increased intercellular CO₂ concentration in the leaves of ‘Newhall’ plants, irrespective of the rootstock used.     

Effect on Resistance to Tylenchulus semipenetrans of Hybrid Citrus Rootstocks Subjected to Continuous Exposure to High Population Densities of the Nematode

Experiments simulating interplanting of resistant rootstocks with susceptible rootstocks that maintain high population densities of Tylenchulus semipenetrans in field soil were carried out in microplots at two locations, and in an naturally infested orchard. Selections of Cleopatra mandarin (03) × Poncirus trifoliata (01) 03.01.5 and 03.01.13, Citrus volkameriana (23) × P. trifoliata 23.01.17, Troyer citrange (02) × Cleopatra mandarin 02.03.24, Troyer citrange × Common mandarin (04) 02.04.18, King mandarin (05) × P. trifoliata 05.01.7, and Carrizo citrange were exposed to continuous high population densities of a population of the Mediterranean biotype of T. semipenetrans. The selection 23.01.17 retained its resistance in the microplots and in the field (< 1.2% females and eggs per gram fibrous root of those on Carrizo citrange). The selection 03.01.5 also retained its resistance in the microplots at Moncada (< 0.5% females and eggs per gram fibrous root of those on Carrizo citrange) but numbers of females and eggs per gram fibrous root were 27% and 22% at Amposta, and 139% and 18% in the orchard of those on Carrizo citrange, respectively. The selection 05.01.7 supported equal number of females and 43% eggs per gram fibrous root of those on Carrizo citrange in the nematode-infested orchard. The remaining selections supported high populations of T. semipenetrans.     

Vesicular-arbuscular mycorrhizae screened for Troyer citrange

Summary Troyer citrange is an important citrus root-stock but a slow grower. Eighteen different vesicular-arbuscular mycorrhizal (VAM) fungi were screened for their symbiotic response with Troyer citrange in unsterilized soil. Of the 18 fungi,Glomus macrocarpum (Ruakura),G. caledonicum (Nedlands),G. velum (Nedlands),G. monosporum (Nedlands),Gigaspora margarita (Ruakura), gave the greatest improvements in growth and nutrition, resulting in larger leaf area, plant height, stem diameter, and plant biomass, with higher P, Zn and Cu contents. All the five fungi were statistically on a par with each other in improving the growth and nutrition of Troyer citrange. Inoculations with these fungi allowed the rootstock to reach a stem diameter of pencil thickness, i.e., ready for budding, in about 13–14 months in contrast to 18–19 months without mycorrhizal inoculations, thus saving about 5–6 months.     

硼对两种不同砧木‘纽荷尔’脐橙叶片硼形态影响的差异

 【目的】阐明不同硼处理下枳橙砧木与枳壳砧木‘纽荷尔’脐橙细胞水平上的硼利用能力差异。【方法】采用水培试验,通过测定叶片中不同形态硼(自由态硼、半束缚态硼、束缚态硼)含量并分析其相互关系,研究两种砧木脐橙叶片硼形态的差异。【结果】在缺硼B0(B 0 mg?L-1)、正常硼B05 (B 0.5 mg?L-1)、高硼B50 (B 5.0 mg?L-1)条件下,枳橙砧木的自由态硼含量相对较低,半束缚态硼含量相对较高,枳壳砧木的正好相反;枳橙砧木在B0、B05、B50条件下的R值(半束缚态硼/自由态硼)分别为2.31、1.23和1.06,都远大于相同处理下枳壳砧木的1.80、0.57和0.82;高硼(B 5.0 mg?L-1)条件下,枳橙砧木的自由态硼和半束缚态硼分别为16.39、17.38 mg?kg-1, 都远大于枳壳砧木的8.60和7.02 mg?kg-1;对于束缚态硼,枳橙砧木的脐橙正常供硼与高硼条件下分别为7.34、7.43 mg?kg-1, 已没有显著差异,而枳壳砧木的则是高硼条件下最高,为9.20 mg?kg-1。【结论】与枳壳砧木相比,枳橙砧木脐橙体内硼的跨膜能力更强,细胞水平上的硼利用能力较高,在相对较低硼水平下就能满足细胞壁的需求。     

转基因枳橙中rolABC基因荧光定量表达分析方法的建立

以转rolABC基因枳橙B、D、E系及野生植株为材料,制备标准品,采用SYBR Green I染料,构建rol基因和β-actin基因的双标准曲线,校正引物扩增效率后,用2-△△Ct法对rol基因在嫩茎、嫩叶、功能叶、树皮和根中的mRNA水平进行相对定量,建立适合于rolA、rolB、rolC基因实时荧光定量RT-PCR分析方法.初步认为rolC基因在嫩茎、嫩叶、功能叶和树皮中表达量最高;其次是rolA基因,主要在嫩茎中表达;rolB基因表达量最低,主要在根系中表达.