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標題: 花粉活力、貯藏及授粉於辣椒採種之研究
Studies on Pollen Viability, Preservation and Pollination in Seed Production of Hot Pepper (Capsicum annuum L.)
作者: 郭宏遠
Kuo, Horng-Yeuan
關鍵字: 花粉活力
pollen viability
pollen germination
desiccation tolerance
heat tolerance
male sterile line
nectar secretion
出版社: 園藝學系所
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摘要: 辣椒為世界性重要之辛香類蔬菜,因為主要品種均為雜交一代品種,因此維持品種純度及種子品質十分重要。由於氣候暖化及採種成本之問題,對於耐熱品系、F1品種親本保護及降低採種成本之需求殷切。本研究之目的擬先進行辣椒耐熱性品系之篩選、花粉品質測定、花粉液態氮保存及評估利用蜜蜂授粉採種之可行性。本研究試驗結果顯示利用離體發芽法(10 %蔗糖濃度的B&K固體培養基)及化學染色法(0.5 % TTC溶液)均能快速檢測花粉活力。以生育期之各項花粉表現從參試的四個品系中篩選出hp27為相對耐熱之品系,供後續設施內試驗之用。hp27的花粉發芽率與花苞長度具正相關性,可當作田間快速判斷花粉活力之指標。植株定植2週後,每週以不含硼肥(B)之Hoagland Solution及2.5 ppm B灌注土壤一次,可以提升花粉之活力與種子品質及增加種子數量。 花藥相對含水量會影響花粉之活力,本研究以四種乾燥條件進行辣椒花藥乾燥,在花苞預乾燥一天或兩天之結果顯示花苞預乾燥二天時,以氯化鈣或低溫乾燥箱(15℃/ 8 % RH)處理之花粉發芽率最高,分別為81.2 %及81.0 %。花苞乾燥過程中,花粉含水量下降、澱粉含量下降及蔗糖含量增加。 為瞭解辣椒花粉保存之適溫,以應用於種原保存、育種及採種。本研究採不同溫度及液態氮進行花粉保存,試驗結果顯示室溫下花粉只能維持3天活力,在4 ℃下9週後已無發芽能力。於-20 ℃及液態氮保存12週後,對授粉後之單果種子數及種子發芽率均無影響。液態氮貯藏花粉後之回溫條件以40 ℃水浴回溫處理3-5分鐘為宜。花粉經液態氮貯藏1年後仍能保持其活力、構造及遺傳穩定性,在超低溫下誘導花粉產生新蛋白質(fructokinase-2)及花粉發芽時之ATP含量較高。 本研究以網室內栽培之辣椒雄不稔品系評估應用蜜蜂授粉於辣椒採種上之可能性,試驗結果顯示,雄不稔品系C在六個參試品系中的花朵大小、花蜜分泌量及吸引蜜蜂訪花頻率均較大,為適合在密閉網室內利用蜜蜂授粉採種之品系。在網室內進行蜜蜂授粉採種時,以父母本(父本:hp27,母本:C)比例1:1於同一畦內交錯定植,可得較佳之採種量。利用蜜蜂攜粉授粉時,以新鮮或液態氮貯藏之hp27花粉均能成功著果及結籽,但少於以人工授粉者。 綜合上述研究結果顯示,hp27為相對耐熱之品系,在植株定植2週後,每週以土壤灌注不含硼肥(B)之Hoagland Solution及2.5 ppm B一次,可以提升花粉之活力。採收開花前一天之花苞進行乾燥處理兩天,待花藥相對含水量降至45-50 %時,再採收花粉進入液態氮貯藏,回溫條件為40 ℃水浴3-5分鐘。採種時以hp27為父本,雄不稔品系C為母本,於網室內利用西洋蜜蜂進行省工授粉及攜粉授粉均能成功,唯採種量低於人工授粉,減少約43.5 %。
Hot pepper (Capsicum annuum L.) is the important spicy vegetable in the world. For commercial F1 cultivars, the variety purity and seed quality is considered to be very important. Due to the climate warming and plant variety protection, the demand of heat-tolerant lines, protection of the parental lines of hybrid variety, cut-down the cost of seed production grow up increasingly. The purpose of this study were to select the heat-tolerance lines, test the pollen quality, and storage pollen in liquid nitrogen, providing for assessing the feasibility of honeybees to carry pollen for pollination. The results showed that in vitro method (pollen incubated on B&K solid medium with 10 % sugar) and histochemical method ( 0.5 % TTC solution) were both rapid test methods for pollen vigor. Line hp27, having better heat-tolerance, was selected from four entry lines by pollen performances through the growth period. There was a positive correlation between pollen germination percentage and flower bud length, which could be used as a rapid test method for pollen vigor in the field. It was helpful for enhancing pollen vigor, seed number and quality by soil irrigated Hoagland Solution(-B) plus 2.5ppm B once a week after transplanting into the pot for two weeks. The relative water content in anther would affect pollen vigor. The effects of the different dehydration treatments on anther relative water content and pollen viability of hot pepper was studied. The results of the flower bud predehydration for one and two days showed that the pollen germination percentage dried by calcium chloride and desiccators (15℃/ 8 % RH) were 81.2 % and 81.0 %, respectively. The decrease of pollen moisture content and starch as well as the increase of sucrose were occurred during flower bud dehydration process. In order to understand the suitable temperature for pollen preservation of hot peper, different temperatures and liquid nitrogen were used as storage conditions to apply on germplasm preservation, breeding and seed production of hot pepper were researched in this study. The results showed that pollen could only maintain vigor for three days in room temperature. The vigor of pollen storaged in 4 ℃ consistently declined until ninth week that no germination was obtained. In addition, the seeds per fruit and seed germination percentage of both -20 ℃ and cryopreservation treatments were still the same with fresh pollen treatment. Furthermore, 40 ℃ water bath for 3-5 min was suitable for thawing after pollen cryopreservation. The pollen cryopreserved for one year could maintain its vigor, structure and genetic stability. New protein (fructokinase-2) in pollen was induced by super chilling and higher ATP content was detected in the germinating pollen. The cytoplasmic male sterile lines of hot pepper were grown in the net-house for assessing the feasibility of honeybee pollination in seed production of hot pepper in the study. We found that CMS line C was suitable for honey bee pollination in seed production due to its larger flowers, more nectar secretion, and higher honey bee foraging frequency compared to other lines tested. When relying on honey bees to do pollination in the closed net-house, the male and female parents (♂:hp27,♀:C) grown in 1:1 ratio within the same row could obtain more seed production. Both fresh and liquid nitrogen preserved pollen could be used for honeybees to carry them for pollination and make successful fruit-sets and seed production. However, using bee pollination yielded less seeds than using hand-pollination. After generalized assessment of study results described above, line hp27 had a better heat-tolerance, whose pollen vigor was enhanced by the following fertilizer treatment: Hoagland Solution(-B) plus 2.5ppm B by soil irrigation once a week after transplanting into the pot for two weeks. Pollen should be preserved in the liquid nitrogen when the relative moisture content of the anther was dried to 45-50% after flower bud pre-dehydrated for two days. Both labor-saving and carrying-pollen pollination by honeybee with cytoplasmic male line C grown in the net-house were successful for fruit-sets and seed production, but using bee pollination this way yielded less seeds than using hand-pollination about 43.5 %.
其他識別: U0005-1601201311295600
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