Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/98187
標題: 以熱療法進行'陽光麝香'葡萄組培苗去病毒及催芽劑對葡萄萌芽之影響
Virus Elimination of Plantlet in vitro of 'Shine Muscat' Grape by Thermotherapy and the Effect of Bud Forcing Chemicals on Budbreak of Grapevines
作者: 謝鍹枰
Hsuan-Ping Hsieh
關鍵字: 葡萄
熱療法
去病毒
健康種苗
催芽
催芽劑
Grape
Thermotherapy
Virus Elimination
Health Seedling
Bud Forcing
Chemical Forcing
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摘要: 'Shine Muscat'('陽光麝香'),以母本為Akitsu-21及父本為Hakunan雜交選育,二倍體鮮食葡萄,由日本於2006年進行品種登記,該品種果實外觀呈黃綠色、果粒大、糖度高、酸味少、果皮可食用且具有特殊香氣,可引進台灣作為葡萄產業具推廣潛力之品種。但在栽培過程中其葉片會表現出畸形、捲縮及褪綠斑駁等症狀,可能與受病毒感染有關,因此本研究乃利用組織培養的方法建立瓶苗,以提供熱療法並配合莖頂組織培養進行去病毒處理,將獲得之無特定病毒苗供大量繁殖及生產健康種苗使用,並利用不刻傷塗抹催芽劑之方式找出適合'Shine Muscat'葡萄商業生產的方法。 採取溫室與田間之腋芽培植體,利用1% NaOCl消毒7、8、9、10及15分鐘後,植入培養基中培養30天,採取溫室之培植體以消毒10分鐘結果顯示之成活率最高、汙染率最低,而採取室外之培植體消毒10或15分鐘顯示之成活率及汙染率表現較佳。另外,採取溫室之培植體消毒15分鐘後其褐化數是達到最多。 切取初代培養之枝梢培養於含有0、0.1、1、0.2、2 mg/L IBA之1/2 MS培養基中,30天後以添加2 mg/L IBA之枝梢發根率、根長及根數來得最好,反而以添加0.1 mg/L IBA之發根率最低。 以37°C進行熱療法去病毒處理30、35、40及45天後其莖頂存活率均在90%左右,而植株再生率及每4週繼代存活率則以處理30天最低,可能為處理前培植體只經1次繼代,尚不足恢復幼年性,導致經熱逆境後莖頂褐化程度較嚴重。經不同處理天數後,組培苗之GLRaV-3去病率均為100%。 以不刻傷塗抹2-氯乙醇、氰胺及蒸餾水進行催芽,其中以2-氯乙醇催芽之萌芽率高、萌芽整齊且新梢長度一致,而氰胺催芽之萌芽雖較早,但萌芽率較2-氯乙醇處理低,可能為氰胺可使Ca2+-ATPase被活化,在解除休眠上扮演訊息傳遞之作用,然而在'Shine Muscat'枝條中Ca及B濃度較'Kyoho'及'Honey Red'來得低,推論'Shine Muscat'可能為易缺Ca之品種。無花穗之'Shine Muscat'葉片中錳元素濃度及鈣硼比顯著高於盛花時之'Shine Muscat',推測為花穗發育時與新梢上葉片之生長存在營養競爭之關係,植物體內來不及進行分配運移,導致生長期之'Shine Muscat'植株發生葉片畸型、捲縮等症狀之原因。
'Shine Muscat' is a diploid table grape cultivar which is a hybrid between Akitsu-21 and Hakunan. It was registered as No.13,891 under the Seed and Seedlings Law of Japan in 2006. It has large yellow-green berries, crisp flesh texture, muscat flavor, high soluble solids concentration and low acidity, which make them to be popular in Taiwan. However, the plant displayed transplanting leaf deformation, leaf curl and chlorotic mottle during cultivating in field which may be due to virus infection. Thus, the objectives of this study were utilized plant tissue culture to establish plantlet in vitro which offer thermotherapy combined with shoot tip culture to eliminate of virus-infected plant. After treatment, the specific virus-free health seedling can be derived, exploited for rapid multiplication and produced health seedling of this plant cultivar. Furthermore, the use of chemical forcing without scarification methods to reaserch in best commercial production of 'Shine Muscat' Grape. The axillary bud explants took from greenhouse and the field were sterilized in 1% NaOCl for 7、8、9、10 and 15, cultured on medium duration 30 days, the survival percentage was the highest, and the contamination percentage was decreased of the explants in greenhouse were sterilized for 10 min. The survival and contamination percentage were showed better of the explants in the field were sterilized for 10 or 15 min. In addition, the browning number was the most of the explants in greenhouse were sterilized for 15 min. Dissected new shoots were cultured on the half strength of MS medium contains 0、0.1、1、0.2、2 mg/L IBA, supplemented with 2 mg/L IBA were getting better of rooting percentage、root number and root length on the 30th day after culture. Instead of supplemented with 0.1 mg/L IBA were getting decreased of rooting percentage. A high survival rate of in vitro-shoot tip of 90% was achieved by thermotherapy at 37°C for 30、35、40 and 45 days, regeneration efficacy and survival rate of subculture every four weeks were decreased of the explants were treatment for 30 days.It is presumed that the explants were subcultured only one time before treatment, have not recover its enough juvenility yet, and caused browning degree of shoot tip much severity after heat stress. The result of 100% GLRaV-3 virus elimination of plantlet in vitro after different treatment day. The use of 2-chloroethanol、hydrogen cyanamide、distilled water without scarification methods to carry out bud forcing, the budbreak was the highest, budbreak uniformity, and the shoot length did not very significantly by 2-chloroethanol. Besides, the time of budbreak was the earliest of hydrogen cyanamide, but the budbreak was lower than 2-chloroethanol treatment. It is presumed that hydrogen cyanamide was able to activate Ca2+-ATPase, and calcium was played a role in signal transduction on release dormancy. Furthermore, the concentrations of calcium and boron on 'Shine Muscat' canes were less than 'Kyoho' and 'Honey Red', maybe 'Shine Muscat' is easy cultivar to absence of calcium. The concentrations of manganese and calcium/boron ratio on 'Shine Muscat' leaves without inflorescence were significantly higher than with inflorescence. It is presumed that there was a nutrient-competed relationship between the development of inflorescence and the growth of leaf in new shoot, no time enough be carried out to distribute or transport, and caused 'Shine Muscat' plant displayed leaf deformation、leaf curl and chlorotic mottle during growing season.
URI: http://hdl.handle.net/11455/98187
文章公開時間: 2021-08-07
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