拮抗酵母對(duì)柑橘采后病害的防治效果及其作用機(jī)理初探

1、1西藏拮抗酵母對(duì)柑橘采后病害的防治效果及其作用機(jī)理初探EvaluationofAntagonisticYeastfromTibetagainstPostharvestDecayofCitrusitsMechanism摘要摘要從西藏地區(qū)分離和篩選對(duì)柑橘主要采后病害綠霉病和青霉病具有良好生防治果的耐低溫拮抗酵母。其中一株效果顯著，經(jīng)鑒定為羅倫隱球酵母（Cryptococcuslaurentii）。當(dāng)該酵母細(xì)胞懸浮液濃度為108個(gè)mL時(shí)，在4

2、℃下分別能抑制柑橘綠霉病和青霉病發(fā)病率至30%和20%，而相同條件下對(duì)照的發(fā)病率為100%。該酵母不同處理液的體內(nèi)試驗(yàn)和體外孢子萌發(fā)試驗(yàn)結(jié)果表明其主要作用機(jī)理仍為與病原菌的營(yíng)養(yǎng)和空間的競(jìng)爭(zhēng)作用。而其分泌的生防相關(guān)酶活性較高，是其低溫生防效果良好的另一主要原因。該酵母胞內(nèi)高含量的海藻糖和脯氨酸使其對(duì)低溫的耐受性增強(qiáng)，從而在低溫條件下能夠更好地抑制水果病原菌的生長(zhǎng)，達(dá)到防腐保鮮效果。同時(shí)本論文也證明了從高寒地區(qū)獲得耐低溫拮抗酵母的可能性。關(guān)

3、鍵詞關(guān)鍵詞:生物防治、采后、羅倫隱球酵母、西藏、柑橘AbstractColdadaptedyeastswereisolatedfromTibetevaluatedasapotentialbiocontrolagentsagainstgreenmold(Penicilliumdigitatum)bluemold(Penicilliumitalicum)ofcitrusfruitincoldstage.Onestrainoftheyeast

4、sanisolateidentifiedasCryptococcuslaurentiiwasfoundtoexhibitthegreatestbiocontrolactivityamongthedifferentisolatesthatwerescreened.Aconcentrationof108cellsmLofthestrainreducedtheincidenceofdecayofgreenmoldbluemoldto30%20

5、%incoldstagerespectivelycomparedtothecontrolwheredecayincidencewas100%.Invivoinvitroexperimentsoftheyeastindicatedthatits3目錄摘要..............................................................................................

6、....................................................1Abstract.............................................................................................................................................12實(shí)驗(yàn)材料與方法.........

7、.................................................................................................................62.1主要儀器...................................................................................................

8、....................62.2試劑...............................................................................................................................62.3受試果實(shí)...........................................................

9、............................................................72.4病原菌...........................................................................................................................72.5拮抗酵母......................

10、.................................................................................................82.6西藏拮抗酵母在柑橘果實(shí)體內(nèi)復(fù)篩試驗(yàn)....................................................................82.7菌種鑒定..........................

11、.............................................................................................92.8西藏羅倫隱球酵母在不同溫度條件下的防腐保鮮效果............................................92.9西藏羅倫隱球酵母不同處理液對(duì)柑橘的防腐保鮮效果.............................

12、.............102.10西藏羅倫隱球酵母對(duì)病原菌孢子萌發(fā)的抑制效果................................................112.11拮抗酵母生防相關(guān)酶活測(cè)定...................................................................................112.11.1酶的提取...................

13、......................................................................................112.11.2幾丁質(zhì)酶活性的測(cè)定.....................................................................................112.11.3?13葡聚糖酶活性的測(cè)定............

14、.............................................................122.12拮抗酵母耐低溫物質(zhì)含量測(cè)定...............................................................................122.12.1樣品制備.................................................

15、..........................................................122.12.2海藻糖含量的測(cè)定...........................................................................................122.12.3脯氨酸含量的測(cè)定.......................................

16、....................................................122.13數(shù)據(jù)處理與分析.......................................................................................................133結(jié)果與分析............................................

17、....................................................................................133.1西藏拮抗酵母在柑橘果實(shí)體內(nèi)復(fù)篩試驗(yàn)...................................................................133.2菌種鑒定......................................

18、...............................................................................143.3西藏羅倫隱球酵母在不同溫度條件下的防腐保鮮效果...........................................143.4西藏羅倫隱球酵母不同處理液對(duì)柑橘的防腐保鮮效果..........................................

19、.183.5西藏羅倫隱球酵母對(duì)病原菌孢子萌發(fā)的抑制效果...................................................223.6拮抗酵母生防相關(guān)酶活測(cè)定......................................................................................243.7拮抗酵母耐低溫物質(zhì)含量測(cè)定.....................