In vitro induction, regeneration and analysis of autotetraploids derived from protoplasts and callus treated with colchicine in Citrus

In the present paper attempts were made to induce chromosome doubling of ‘Meiwa’ kumquat (Fortunella crassifolia) protoplasts and ‘Frost’ navel orange (Citrus sinensis Osbeck) embryogenic callus via colchicine treatment. Colchicine decreased protoplast viability, delayed protoplast division and inhibited callus growth, indicating presence of toxicity to cells. Cell lines established from ‘Meiwa’ protoplasts treated with 0.01 and 0.1% colchicine for 8, 16 and 24 h at each concentration showed different responses when they were cultured on embryoid-induction medium. Flow cytometry (FCM) demonstrated that tetraploids were detected in cell lines and embryoids from all of the treatments, with the highest frequency being 19.23%. As for ‘Frost’, tetraploid cells were only detected when the callus was treated with 0.1% colchicine for either 4 or 8 days, from which plantlets were regenerated. FCM and chromosome counting confirmed them as true tetraploids. The diploid cells were more active in mitotic division during a 12-day culture and smaller in size than their tetraploid counterpart. Potential applications of the novel tetraploid germplasms obtained through in vitro chromosome doubling to citrus cultivar improvement are discussed.     

湖北省纽荷尔脐橙果实品质状况的研究

以纽荷尔脐橙为试材,分析了湖北省13个产地果实的常规品质和果皮色差值。结果显示,不同产地纽荷尔脐橙果实品质差异明显。系统聚类结果表明,秭归县、巴东县和兴山县的常规品质聚为第一类,长阳县、郧县、公安县、枝江市和宜都市聚为第二类,而丹江口市、阳新县、通城县、天门市和京山县聚为第三类;就果皮色差值来看,长阳县、郧县、兴山县和宜都市聚为第一类,巴东县、京山县、枝江市、丹江口市、阳新县、公安县、秭归县聚为第二类,天门市和通城县聚为第三类。综合分析常规品质和果皮色差值可以看出,三峡库区的兴山县、秭归县、巴东县纽荷尔脐橙综合品质最好,邻近三峡库区的长阳县、宜都市、枝江市,丹江库区的郧县和卷桥库区的公安县次之,丹江口市、阳新县、京山县,尤其是天门市和通城县的品质表现不如上述各个产地。因此湖北省纽荷尔脐橙应重点集中在三峡库区这一优势区域内优先发展,同时向其邻近的适宜地区辐射,而在其它产地宜选择小气候适宜地区适当发展。     

Stable maintenance and expression of a foreign gene in transgenic pear shoots retrieved from in vitro conservation

Shoot-tips of transgenic pear were successfully conserved in vitro by slow-growth and cryopreservation methods. After 1 year of storage in slow growth conditions, all shoot-tips survived showing shoot re-growth. Similarly, shoot-tips showed high survival and regeneration rates after cryopreservation. The genetic stability of the transgenic GUS construct in shoots recovered from slow-growth and cryopreservation was assessed. The results from polymerase chain reaction and Southern blotting showed the stable maintenance of the GUS gene in genome, and the Single-Strand Conformation Polymorphism assay did not detect single base variation. X-Gluc staining suggested a normal expression of the GUS gene in shoots retrieved from these storage methods.     

脐橙晚熟突变体"奉晚"与原品种"奉节72-1"的果实着色差异

测定了果实成熟过程中“奉晚”和“奉节72．1”两个脐橙（Citrus sinensis L．Osbeck）品种果皮中叶绿素和总类胡萝卜素的含量,并采用RT-PCR技术研究了相关酶的基因表达。结果表明：“奉晚”脐橙果皮中叶绿素含量的显著降低发生在11月份,“奉节72．1”则发生存10月中下旬到11月上旬;“奉晚”脐橙果皮中的总类胡萝卜素含量显著积累始于12月中旬,而“奉节72.1”品种则始于11月初。另外,“奉晚”脐橙果皮中叶绿素含量在10～11月显著高于原品种,总类胡萝卜素含量在12-1月显著低于原品种。从基因表达分析结果看到,“奉晚”脐橙果皮中与类胡萝卜素合成酶相关的基因较强表达的时间也整体迟于“奉节72．1”脐橙;叶绿素水解酶基因表达在10～12月中旬表达较“奉节72—1”弱,翌年1月份则较“奉节72．1”强。     

Pilot testing ofKloeckera apiculata for the biological control of postharvest diseases of citrus

The efficacy of the yeastKloeckera apiculata, strain 34-9, in controlling postharvest decay of citrus fruit was evaluated in small-scale and pilot tests in commercial packinghouse. Kloeckera apiculata grew efficiently on different media and maintained its antagonistic activity against spore germination ofPenicillium italicum. In small-scale experiments with citrus fruits dipped in the yeast cell suspension, the development of decay in citrus was effectively inhibited. The yeast was compatible with a mixture of low concentration of a commonly chemical fungicide. In packinghouse tests, combining the yeast with 40 mg/kg Carbendazim (MBC) resulted a reduction in the incidence of decay to a level equal to that of the commercial treatment of 200 mg/kg MBC. The efficacy of the strain 34-9 could also be maintained under packinghouse conditions at a cell concentration of the yeast antagonist as low as 10⁶ cells/ml. No significant difference in the efficacy ofK. apiculata was found in either the drench or the spray application systems tested in citrus packinghouse. Scanning electron microscopy revealed attachment of the yeast cells to the pathogen hyphae. The high antagonistic activity of strain 34-9 against citrus blue mould may be related to its capability to compete withPenicillium italicum, for space and nutrients and /or involvement of directly antagonist of the yeast on the fungus.