A novel method for Citrus propagation: Seed grafting

Summary

We have developed a novel technique for grafting citrus seeds onto citrus rootstock plants that resulted in successful graft-take with normal vascular connections between the emerging seedling stem tissues and the rootstock plant. The method was found to be suitable for producing grafted plants from seeds of six cultivars and hybrids of Citrus and the citrus relative Murraya paniculata, using four common Citrus rootstocks. Plants produced by this method developed normally and were established in the field more rapidly than those produced by the common practice of grafting the rootstocks with budwood derived from seedlings prepared from seed in soil-based media. Seed grafting is expected to find a range of uses in breeding programmes; for example, by reducing the time required for the evaluation of hybrid seedlings, in cases where the female parent is mono-embryonic, for testing for vertical transmission of pathogens, and for screening for pathogen resistance among hybrid and mutagenised seed sources.     

两个柑桔杂交组合F_1代的几种同工酶的遗传分析

采用聚丙烯酰胺凝胶电泳对柑桔两个杂交组合克里迈丁×三湖化红、克里迈丁×宜昌橙F_1代杂种各30侏的GOT、SOD、POX、ME四种酶的同工酶进行了检测分析,发现在GOT—2、SOD—2、POX—2、ME—2四个酶位点上F_1代有分离,表明亲本在这些酶基因位点上处于杂合状态。GOT—2、SOD—2、POX—2的分离经x~2测验,符合孟德尔单基因遗传,与其相应的Got—2、Sod—2、Pox—2三个酶基因可作为遗传标记应用。ME—2的分离复杂,说明柑桔在遗传基础上十分复杂。     

Olive seedling first-flowering position and management

The prolonged length of the juvenile period represents a substantial obstacle in olive (Olea europaea) breeding programs, delaying both the possibility for analyzing the fruit, the harvest unit, and the capacity for sexual recombination. In both olive and other fruit tree species, the juvenile–adult transition has been successfully hastened by forcing and formation procedures designed to rapidly achieve a minimum height. Precise knowledge of the position within the tree canopy where the juvenile–adult transition occurs, identified by the location of the first flowers, offers further potential for manipulating tree structure in order to hasten that transition. The occurrence of the juvenile–adult transition has been described spatially within the canopy as the formation of a juvenility cone, which we report here for olive plants from seed, and for the first time quantitatively describe its position based on the added trunk and branch distances from the soil. In canopies of seedlings from open pollination of cv. Arbequina olive the first-flowering position was consistently located, in our conditions, at an average distance of 200 cm from the trunk base. Based on that evidence, that first flowering can occur at lower positions, and on the values we obtained, three canopy-formation heights (100–130 cm, 130–160 cm and >160 cm) were compared for their effect on the length of the juvenile period and the vigour of olive seedlings. Canopy heights of 100–130 cm, much lower than the 160 cm previously reported by the olive tree breeding program of Córdoba, produced the highest number of flowering plants in the first two flowering years and also provided easier management.     

世界柑橘品种改良的进展

 柑橘鲜食品种的发展趋势是容易剥皮、无核、有香味和风味浓。过去30年, 通过芽变选种、杂交育种等途径, 全世界培育出了上百个柑橘品种。芽变选种获得的新品种主要来自如温州蜜柑、脐橙和克里曼丁橘, 它们分别形成了品种群, 不同品种采收期可长达4～6个月; 杂交育种途径培育出70多个品种(系) , 主要利用了单胚性品种克里曼丁橘、胞质雄性不育的温州蜜柑以及外观色泽好的红橘等作为杂交亲本; 新的砧木品种(系) 主要来自中国的枳与酸橘等品种的杂交; 加工品种的选育相对落后。生物技术, 特别是胚抢救及细胞融合等技术大大加快了柑橘育种步伐。     

板栗品种雌雄异熟的生物学特性

3个板栗区域性试验点,16个品种连续4年的物候观察和主要经济性状调查表明:板栗品种表现出较强的雌雄异熟性(81.25%)。根据花期物候,可将板栗品种分为雌先型、雄先型、相遇型。雌雄异熟秩序不受生态、气候条件、年份的影响。雌雄异熟性与产量、嫁接后2～3年结果株率等性状没有必然联系,而与坐果率显著相关。弄清板栗品种的雌雄异熟性对杂交育种、生产中授粉品种的配置有指导意义,建议在今后的育种、栽培中对这一特殊物候性状加以重视。     

Cold treatment modifies the photoperiodic flowering response of Lobelia×speciosa

Lobelia×speciosa Sweet ‘Compliment Scarlet' was grown under a range of photoperiods and low temperature treatments to determine their effects on flowering. In the first experiment, plants were held at 5°C for 0 or 15 weeks, then grown at 20°C under the following photoperiods: 10, 12, 14, 16, or 24 h of continual light or 9 h with a 4 h night interruption (NI). Non-cooled ‘Compliment Scarlet' flowered as a qualitative long-day plant (LDP) with a minimum flowering photoperiod of 14 h. Following cold, flowering was quantitative with respect to photoperiod, until ≈14.2 h, when the calculated rate of progress toward flowering reached a plateau. In cooled plants, node number below the inflorescence decreased from 27 to 16 as the photoperiod increased from 10 to 24 h. Cooled plants developed 61–149% more flowers and were ≥17% taller than non-cooled ones under the same photoperiod. To determine the cold duration required for flowering under short days (SD), plants were held at 0, 3, 6, 9, 12, or 15 weeks at 5°C then grown at 20°C under SD (9 h photoperiod) or long days (9 h photoperiod with a 4 h NI). Under SD, few plants flowered after ≤6 weeks of cold. As cold treatment increased from 9 to 15 weeks, flowering percentage increased, time to flower decreased from 93 to 64 days, and node count decreased from 24 to 13. Cold treatment did not affect flowering percentage or time under NI, but plants always had more flowers and were taller than reproductive ones under 9 h day lengths. Thus, ‘Compliment Scarlet', is a qualitative LDP, but an extended cold treatment can partially substitute for the long day (LD) photoperiodic requirement.     

根癌农杆菌介导的CG1-400-RNase基因转化创制锦橙转基因再生植株

【目的】为了快速有效地利用基因工程的方法获得柑橘无核新种质,【方法】以我国特色多核柑橘优良品种锦橙实生苗上胚轴切段为外植体,采用根癌农杆菌介导法进行能导致种子败育基因CG1-400-RNase的转化;为快速有效地筛选出转化子,在实生苗上胚轴切段转化再生过程中,根据不同发育阶段的组织或器官对抗生素的敏感程度不同采用不同的选择压。【结果】结果表明,在抗性芽再生过程中卡那霉素质量浓度设定为50 mg.L-1,获得的362个抗性芽转入卡那霉素质量浓度为100 mg.L-1的伸长培养基中进行伸长培养后,进行早期PCR检测,获得28个阳性芽;经过不定芽诱导生根或试管嫁接,获得22株完整植株。【结论】再生植株经PCR和Southern杂交检测,获得2株目的基因以单拷贝的形式插入锦橙基因组的转基因植株,为最终获得具有无核性状且可稳定遗传的柑橘新种质奠定了基础。     

Beneficial roles of arbuscular mycorrhizas in citrus seedlings at temperature stress

Citrus, a cold-sensitive plant, often suffers from low temperature, which seriously affects citrus productivity. The objective of the study was to elevate the roles of an arbuscular mycorrhizal fungus, Glomus mosseae, in growth, photosynthesis, root morphology and nutrient uptake of citrus (Citrus tangerine) seedlings under temperature stress conditions. Three-month-old seedlings with or without G. mosseae were grown for 55 days at moderate temperature (25 °C) and low temperature (15 °C). Low temperature severely restrained symbiotic development including mycorrhizal colonization, entry point, vesicle and arbuscule relative to moderate temperature. Mycorrhizal seedlings grown at 25 °C maintained better stem diameter, plant height, leaf area, root and total dry weights, higher photosynthetic rate, transpiration rate and stomatal conductance, higher root volume, and more uptake of P, Ca and Mg relative to corresponding non-mycorrhizal control. However, mycorrhizal inoculation significantly increased only the root length and the Ca content of the seedlings grown at 15 °C. The results indicated that mycorrhizal formation had the beneficial effects on growth, photosynthesis, root morphology and part nutrient uptake of citrus seedlings grown at moderate temperature, but the beneficial roles of arbuscular mycorrhizas were almost lost at low temperature.     

乐东拟单性木兰绿化苗木繁育技术及生长规律的研究

对乐东拟单性木兰播种苗及2 a生以上绿化苗木进行培育技术和生长规律研究。结果表明:乐东拟单性木兰大田育苗播种量以20～25 g/m~2为宜。1 a生苗高平均55.6 cm,平均地径0.67 cm,主根长26.1 cm,根冠32.6 cm×29.9 cm,5 cm长Ⅰ级侧根数9.8根,根系较发达,当年可出圃。10 a生绿化大苗平均高达(617±31.97)cm,平均胸径(15.66±1.18)cm,冠径(447±31.85)cm,生长速度较快。每年的6～8月是苗高、径生长最快时期,应加强肥水管理,促进苗木生长。     

Growth,root morphology and boron uptake by citrus rootstock seedlings differing in boron-deficiency responses

Boron (B) is an essential microelement for higher plants and has important physiological functions in plant growth and development. Citrus plants are frequently exposed to B-deficiency, but knowledge regarding the effects of B-deficiency on rootstock growth, root morphology and genotypic variations in citrus is limited. To evaluate the variations in plant-growth parameters in response to B-deficiency, five citrus rootstocks seedlings that grown in modified 1/2-strength Hoagland's solution that contained 0.25 μM H₃BO₃ (moderate B-level, Control) or 0 μM H₃BO₃ (B-deficiency, −B) were investigated before and 90 days after treatment. There are significant genotypic variations in plant-growth parameters (dry mass, leaf area and seedling height), root-morphological traits and B-concentrations. B-deficiency inhibited plant dry mass and leaf area expansion of Fragrant citrus, Sour orange and Trifoliate orange seedlings significantly (P < 0.05), whereas no effect was found on the total dry mass of Carrizo citrange and Red tangerine seedlings. Further, B-deficiency reduced the root number, root length and root surface area in Fragrant citrus and Trifoliate orange significantly (P < 0.05); however, no significant effect was detected on these root morphological traits of Carrizo citrange and Red tangerine (P > 0.05). In addition, B-deficiency reduced the B-concentration in the leaves of all the five genotypes and the B-concentration in the roots of Fragrant citrus and Sour orange, none effect was observed on the B-concentration in stems of all studied genotypes and roots. However, there were significant relationships between the root-morphological traits and B-uptake efficiency. These results showed that Carrizo citrange and Red tangerine are B-efficient, Trifoliate orange is the moderate B-efficient genotypic rootstocks, whereas Fragrant citrus and Sour orange are B-inefficient genotypic rootstocks. In addition, Trifoliate orange was the moderate B-efficient rootstocks of five genotypes. The root-to-shoot dry mass ratio (R/S) and longer root numbers were higher in the Carrizo citrange and Red tangerine than that of Trifoliate orange, Fragrant citrus and Sour orange genotypes. That is to say, maintaining higher R/S and numerous longer lateral roots is very important in improving the B-efficiency and thus contribute much to the resistance of seedlings to B-deficiency in the Carrizo citrange and Red tangerine genotypes.     

梨杂种后代亲本性状遗传倾向的研究

通过对49个杂交组合2542株梨杂种实生苗亲本性状遗传倾向的观察研究看出,在河南郑州地区,梨杂种后代的童期为4～5年,6年生杂种一般开花株率可达70%以上;童期的长短主要受亲本遗传特性的制约;梨杂种后代果实重量一般较亲中值小27%,但有些组合超亲现象明显,一般超亲19.2%;果形遗传似表现为圆或扁圆对卵圆或圆锥为显性;皮色遗传表现为黄对褐为显性;肉质的遗传表现为细脆、紧脆和粗脆对细嫩占优势,细嫩型所占比重约为30%;在品质遗传方面,鸭梨、麻梨、库尔勒香梨易将其优良品质传给后代,在鸭梨×金花梨的后代中,果实大小和风味品质超亲现象明显。     

树莓杂交后代实生苗部分性状的遗传变异分析

对树莓4个杂交或自交得到的197株2a生实生苗的重要经济性状进行了调查和遗传分析.结果表明,亲本性状各异树莓杂交后代表现了相近的果形分布比例,均以圆锥形为主,其中3个组合后代中圆锥形果实超过了75%;短圆锥形和球型果比例相近,总和接近25%,后代中还出现了少量比例的畸形果.杂交后代可溶性固形物含量的分布受亲本影响,但变异幅度较宽.平均单果重受亲本影响极大,大果型品种美国22号和图拉米的后代的大果型比例偏多.图拉米是优良的无刺树莓育种亲本,其自交后代出现了21%的无刺植株.     

Studies on the induction of flowering in juvenile Prunus avium L.

Juvenile Prunus avium shoot apices produced flowering shoots after grafting to mature trees irrespective of treatment of the apices in the season before grafting with G A or G A + cytokinin. Scions grown from mature shoot apices grafted to seedling stocks failed to flower, again irrespective of prior hormone treatment. Similar treatment of mature shoot apices with these hormones, or with zeatin, inhibited concurrent floral initiation, but G A or GA + zeatin treatments increased flowering of scions grown from the treated apices after grafting to untreated mature trees. Localized shoot tip or root drench treatment of one or two year old seedlings with (2RS, 3RS)-paclobutrazol failed to induce flowering, but treatment of three or four year old plants did stimulate flowering. Branch or stem girdling or root flooding applied alone or in combination to three year old plants did not affect flowering. Floral initiation by three or four year old plants was inhibited by treatment with GAs. The results were consistent with the presence in seedlings of a root- produced, xylem transported, graft-transmissible inhibitor(s), which control the initiation of otherwise competent meristems, and which could include factors other than GAs. Juvenile meristem competence to flower was not affected by prior GA treatment.     

Production and evaluation of transgenic sweet orange (Citrus sinensis Osbeck) containing bivalent antibacterial peptide genes (Shiva A and Cecropin B) via a novel Agrobacterium-mediated transformation of mature axillary buds

Transgenic plants of Jincheng orange (Citrus sinensis Osbeck) and Newhall navel orange (C. sinensis Osbeck) containing antibacterial peptide genes Shiva A and Cecropin B were successfully obtained by a novel Agrobacterium tumefaciens-mediated transformation of the mature axillary buds. PCR and Southern blot analysis of the transgenic plants verified that the Shiva A and Cecropin B genes were integrated into the citrus genome. The transgenic plants began to blossom and bear fruit in the 2nd year after grafting on trifoliate orange (Poncirus trifoliata Raf) rootstock in greenhouse. Water-soluble extracts from transgenic citrus leaves exhibit in vitro suppressive effects on Xanthomonas axonopodis pv. citri, suggesting that the expressed products of Shiva A and Cecropin B in citrus retain their native antibacterial activities. Artificial inoculation in greenhouse and open field further indicates significantly increased resistance of transgenic plants to X. axonopodis pv. citri when compared with non-transgenic lines. No significant difference was found in the content of total soluble solids, total acidity, reduced sugar content and other fruit characteristics between transgenic and non-transgenic plants. In this present study, 11 transgenic lines were obtained from 40 transgenic lines, showing enhanced resistance to citrus canker disease.     

单胚槾橘×多胚岩溪晚芦槿柑多胚种子的胚分离培养及后代的叶形态和SSR鉴定

继首次报道单胚清见与多胚桠柑杂交产生高比例多胚种子后,在单胚棱橘（Citrus tardiferax Hort．ex Tan）与多胚岩溪晚芦桠柑（Citrus reticulata Blanco）的杂交组合中,又发现高比例多胚种子,其比例一般为30％左右。为探明多胚来源,对多胚进行了分胚组织培养,获得同一种子来源的多胚...     

柑橘转基因研究的现状及展望

从柑橘种及品种、转化方法、导入的目的基因、转化所用的外植体、转基因频率及其影响因素等方面对柑橘转基因的现状进行了综述,并提出了下一步的研究重点。已成功获得转基因植株的柑橘种类有枳、甜橙、酸橙、宽皮柑橘、柚和来檬;柑橘转基因的手段主要是农杆菌介导法,ＰＥＧ介导法和基因枪法也有应用;目前导入柑橘的目的基因主要是与抗病抗虫相关的基因。近年来涉及柑橘糖、类胡萝卜素、有机酸、乙烯、ＧＡ、柠檬苦素、叶绿素、黄酮类及果胶等代谢以及果实发育、成熟和柑橘开花等生理过程的基因均已分离,为通过遗传工程手段调控柑橘果实物质代谢及发育过程奠定了基础。     

柑橘10个品种实生后代多倍体的发掘及SSR鉴定

秋季采摘柑橘成熟果实并剥取种子,将小粒种籽和瘪籽播于MT培养基,待萌发长出2～3片真叶时,取其叶片检测倍性;将饱满种籽播种于露地,对其幼苗首先通过形态筛选获得似多倍体的植株,然后取其叶片用流式细胞仪进行倍性检测;最后对获得的多倍体植株进行SSR分子鉴定。试验结果表明,获得早金甜橙、卡特夏橙、塔罗科血橙、日辉橘、Ortanique橘橙、默科特橘橙、华农本地早橘、朱红橘和Flame葡萄柚等9个品种的四倍体分别为6、1、3、12、3、6、2、1和10株;获得早金甜橙、塔罗科血橙、Itabori甜橙和Ortanique橘橙等4个品种的三倍体分别为1、1、2、4株。用13对多态性SSR引物对胚抢救获得的三倍体和四倍体的来源进行鉴定,发现早金甜橙、Itabori甜橙的三倍体再生植株为异源三倍体;塔罗科血橙、Ortanique橘橙的三倍体可能为同源三倍体;早金甜橙和Ortanique橘橙的四倍体可能为同源四倍体;塔罗科血橙四倍体为异源四倍体。采用37对多态性引物对从饱满种籽中获得的8个品种四倍体的来源进行鉴定,发现均为同源四倍体。这些自然发掘的多倍体资源对柑橘无核育种和相关基础研究具有重要价值。     

苹果杂种后代亲本性状遗传倾向的研究

通过对20个苹果杂交组合的587侏后代本性状的遗传倾向的研究看出,在河南郑州地区,苹果杂种的童期一般为5年,6年生苹果杂种实生苗一般开花株率为25％,7年生可达50％以上;童期的长短主要受亲本的制约;干周15cm为杂种植株童期即将结束或已结束的标志。苹果杂种初次开花结果的部位主要为短果枝,约占60％,大多数杂种具有腋花芽结果习性。苹果杂种的果实一般均小于亲中值,平均较亲中值小38.5％。色泽的遗传表现为红对黄为显性,但在红×红的后代中有相当比例的绿黄果。果实的风味遗传相当复杂,但无论甜×酸或甜×甜,后代中果实风味酸甜的所占比重最大,并且甜×甜的后代中可出现甜的类型。富士和金冠是培育优质新品种的最好亲本;新红星的红色遗传传递力似乎较强。     

硫酸铜乳胶漆制剂对银荆容器播种苗生长的影响

研究了不同质量分数CuSO4乳胶漆制剂处理容器内壁对银荆播种苗生长的影响。结果表明:不同浓度CuSO4乳胶漆制剂对银荆播种苗主根生长具有抑制作用,浓度为270g/LCuSO4处理后的银荆播种苗主根最短(1.125cm),对照组最长(9.5cm);不同浓度CuSO4乳胶漆制剂对容器苗侧根数量、复叶数量、小叶数量、叶面积、茎重、根重及苗高增加均具有显著的促进作用;190g/L CuSO4乳胶漆制剂处理容器内壁能显著抑制银荆1a生播种苗主根生长,提高苗木质量。