獼猴桃是原產于我國的重要果樹。據統計,我國獼猴桃選育品種累計達180余個,栽培總面積400多萬畝,產量超過300萬噸,兩者均穩居世界第一(超過50%),在脫貧攻堅和鄉村振興事業中發揮了重要作用。獼猴桃是淀粉積累型水果,其淀粉含量占干物質的40%左右,是決定風味和貯藏性能關鍵因素。淀粉體是一種非光合的質體形式,是淀粉代謝的重要場所。一般,質體定位的2000-3000個蛋白質中,僅有150個左右是質體自身編碼的蛋白,其他均為通過TOC/TIC等復雜途徑轉入。因此,探討果實淀粉體的生物發生和分化機制就顯得十分重要。
近日,華中農業大學果蔬園藝作物種質創新與利用全國重點實驗室程運江教授/曾云流副教授課題組在The Plant Journal發表了題為The kiwifruit amyloplast proteome (kfALP): a resource to better understand the mechanisms underlying amyloplast biogenesis and differentiation的學術論文,揭示了獼猴桃果實淀粉體生物發生和分化的潛在分子機制。
我們發現黃肉和綠肉獼猴桃果實的淀粉體的生物發生模式相似,都由葉綠體分化而來。然而,其淀粉體分化模式卻存在差異:黃肉獼猴桃果實淀粉體分化為含有大量質體小球的有色體,并伴隨著淀粉顆粒的降解和類囊體膜的消失,而綠肉果實的葉綠體結構一直比較穩定(圖1)。這很好地解釋了滯綠/褪綠表型,并構成了甜度風味形成的結構基礎。
我們創造性地發明了一種基于密度梯度離心方法的獼猴桃果肉淀粉體的分離提純方法,并將之成功的應用到黃/綠肉的不同發育與成熟期質體的分離提純。基于此,我們首先利用Label-free和TMT技術,解析并構建了目前為止最大的淀粉體蛋白譜庫(圖2)。其次,比較蛋白組學分析表明GWD、PWD及BAM等多個參與淀粉代謝的關鍵蛋白,可能參與淀粉顆粒的降解;TOC/TIC以及ATPase蛋白可能參與淀粉體生物形成與分化。更為重要的是,我們發現光合作用和四吡咯途徑相關蛋白在兩種果實淀粉體分化過程中的變化模式存在顯著差異,推測二者可能是影響淀粉體分化的關鍵蛋白。
綜上所述,上述結果重塑了以淀粉體為中心的質體分化網絡,為獼猴桃果實品質提升,提供了重要研究方向。
華中農業大學園藝學博士李昂為論文第一作者,華中農業大學果樹系曾云流副教授為該論文通訊作者,程運江教授、已畢業研究生林加嘉和浙江省農業科學院鄧志平研究員等參與了研究。新西蘭植物與食品研究院Ross Atkinson, Niels J Nieuwenhuizen、Charles Ampomah-Dwamena 等對該研究進行了指導。本研究得到了國家自然科學基金項目面上項目、國家現代農業柑橘產業技術體系、國家重點研發、湖北省重點研發等項目的資助。
曾云流副教授領銜的研究團隊長期從事果實品質調控研究,受聘為國家現代(柑橘)農業產業體系獼猴桃質量安全與加工保鮮崗位、湖北省高層次人才計劃。質體是植物特有的一類亞細胞器,質體分化是采后成熟與衰老的主要表征,比如轉色、淀粉降解和香氣釋放。課題組以獼猴桃、柑橘果實為材料,建立了有色體(JXB, 2011)、造油體(Horticulture Research, 2018)、淀粉體(Plant Journal, 2023a),及質體小球(Plant Journal, 2023b)的高純度分離提純方法。通過研究溫度、光質、乙烯等信號調控質體分化,如淀粉體-有色體分化(Plant Physiol, 2015),解析了果實風味形成機制,在輔助萜烯類香氣育種(Plant Physiol, 2020)、色澤改良(JXB, 2021)等方面取得了重要突破。同時,開發了配套品質提升技術、工藝與裝備,比如金柑果面/獼猴桃果肉快速褪綠技術,獼猴桃果面色斑去除裝備,獼猴桃變溫即食控熟裝備等等,這些裝備已經在主產區推廣應用,取得了良好效益。
【英文摘要】
The biogenesis and differentiation (B&D) of amyloplasts contributes to fruit flavor and color. Here, remodeling of starch granules, thylakoids and plastoglobules was observed during development and ripening in two kiwifruit (Actinidia spp.) cultivars – yellow-fleshed 'Hort16A' and green-fleshed 'Hayward‘。 A protocol was developed to purify starch-containing plastids with a high degree of intactness, and amyloplast B&D was studied using label-free-based quantitative proteomic analyses in both cultivars. Over 3000 amyloplast-localized proteins were identified, of which >98% were quantified and defined as the kfALP (kiwifruit amyloplast proteome)。 The kfALP data were validated by Tandem-Mass-Tag (TMT) labeled proteomics in 'Hort16A’。 Analysis of the proteomic data across development and ripening revealed: 1) a conserved increase in the abundance of proteins participating in starch synthesis/degradation during both amyloplast B&D; 2) up-regulation of proteins for chlorophyll degradation and of plastoglobule-localized proteins associated with chloroplast breakdown and plastoglobule formation during amyloplast differentiation; 3) constitutive expression of proteins involved in ATP supply and protein import during amyloplast B&D. Interestingly, two different pathways of amyloplast B&D were observed in the two cultivars. In 'Hayward‘, significant increases in abundance of photosynthetic- and tetrapyrrole metabolism-related proteins were observed, but the opposite trend was observed in 'Hort16A’。 In conclusion, analysis of the kfALP provides new insights into the potential mechanisms underlying amyloplast B&D with relevance to key fruit quality traits in contrasting kiwifruit cultivars.
原文鏈接:https://onlinelibrary.wiley.com/doi/10.1111/tpj.16611
日期:2024-01-15
近日,華中農業大學果蔬園藝作物種質創新與利用全國重點實驗室程運江教授/曾云流副教授課題組在The Plant Journal發表了題為The kiwifruit amyloplast proteome (kfALP): a resource to better understand the mechanisms underlying amyloplast biogenesis and differentiation的學術論文,揭示了獼猴桃果實淀粉體生物發生和分化的潛在分子機制。
我們發現黃肉和綠肉獼猴桃果實的淀粉體的生物發生模式相似,都由葉綠體分化而來。然而,其淀粉體分化模式卻存在差異:黃肉獼猴桃果實淀粉體分化為含有大量質體小球的有色體,并伴隨著淀粉顆粒的降解和類囊體膜的消失,而綠肉果實的葉綠體結構一直比較穩定(圖1)。這很好地解釋了滯綠/褪綠表型,并構成了甜度風味形成的結構基礎。
我們創造性地發明了一種基于密度梯度離心方法的獼猴桃果肉淀粉體的分離提純方法,并將之成功的應用到黃/綠肉的不同發育與成熟期質體的分離提純。基于此,我們首先利用Label-free和TMT技術,解析并構建了目前為止最大的淀粉體蛋白譜庫(圖2)。其次,比較蛋白組學分析表明GWD、PWD及BAM等多個參與淀粉代謝的關鍵蛋白,可能參與淀粉顆粒的降解;TOC/TIC以及ATPase蛋白可能參與淀粉體生物形成與分化。更為重要的是,我們發現光合作用和四吡咯途徑相關蛋白在兩種果實淀粉體分化過程中的變化模式存在顯著差異,推測二者可能是影響淀粉體分化的關鍵蛋白。
綜上所述,上述結果重塑了以淀粉體為中心的質體分化網絡,為獼猴桃果實品質提升,提供了重要研究方向。
華中農業大學園藝學博士李昂為論文第一作者,華中農業大學果樹系曾云流副教授為該論文通訊作者,程運江教授、已畢業研究生林加嘉和浙江省農業科學院鄧志平研究員等參與了研究。新西蘭植物與食品研究院Ross Atkinson, Niels J Nieuwenhuizen、Charles Ampomah-Dwamena 等對該研究進行了指導。本研究得到了國家自然科學基金項目面上項目、國家現代農業柑橘產業技術體系、國家重點研發、湖北省重點研發等項目的資助。
曾云流副教授領銜的研究團隊長期從事果實品質調控研究,受聘為國家現代(柑橘)農業產業體系獼猴桃質量安全與加工保鮮崗位、湖北省高層次人才計劃。質體是植物特有的一類亞細胞器,質體分化是采后成熟與衰老的主要表征,比如轉色、淀粉降解和香氣釋放。課題組以獼猴桃、柑橘果實為材料,建立了有色體(JXB, 2011)、造油體(Horticulture Research, 2018)、淀粉體(Plant Journal, 2023a),及質體小球(Plant Journal, 2023b)的高純度分離提純方法。通過研究溫度、光質、乙烯等信號調控質體分化,如淀粉體-有色體分化(Plant Physiol, 2015),解析了果實風味形成機制,在輔助萜烯類香氣育種(Plant Physiol, 2020)、色澤改良(JXB, 2021)等方面取得了重要突破。同時,開發了配套品質提升技術、工藝與裝備,比如金柑果面/獼猴桃果肉快速褪綠技術,獼猴桃果面色斑去除裝備,獼猴桃變溫即食控熟裝備等等,這些裝備已經在主產區推廣應用,取得了良好效益。
【英文摘要】
The biogenesis and differentiation (B&D) of amyloplasts contributes to fruit flavor and color. Here, remodeling of starch granules, thylakoids and plastoglobules was observed during development and ripening in two kiwifruit (Actinidia spp.) cultivars – yellow-fleshed 'Hort16A' and green-fleshed 'Hayward‘。 A protocol was developed to purify starch-containing plastids with a high degree of intactness, and amyloplast B&D was studied using label-free-based quantitative proteomic analyses in both cultivars. Over 3000 amyloplast-localized proteins were identified, of which >98% were quantified and defined as the kfALP (kiwifruit amyloplast proteome)。 The kfALP data were validated by Tandem-Mass-Tag (TMT) labeled proteomics in 'Hort16A’。 Analysis of the proteomic data across development and ripening revealed: 1) a conserved increase in the abundance of proteins participating in starch synthesis/degradation during both amyloplast B&D; 2) up-regulation of proteins for chlorophyll degradation and of plastoglobule-localized proteins associated with chloroplast breakdown and plastoglobule formation during amyloplast differentiation; 3) constitutive expression of proteins involved in ATP supply and protein import during amyloplast B&D. Interestingly, two different pathways of amyloplast B&D were observed in the two cultivars. In 'Hayward‘, significant increases in abundance of photosynthetic- and tetrapyrrole metabolism-related proteins were observed, but the opposite trend was observed in 'Hort16A’。 In conclusion, analysis of the kfALP provides new insights into the potential mechanisms underlying amyloplast B&D with relevance to key fruit quality traits in contrasting kiwifruit cultivars.
原文鏈接:https://onlinelibrary.wiley.com/doi/10.1111/tpj.16611
日期:2024-01-15