








摘""要:甜瓜為海南省重要經濟作物,授粉以激素授粉為主,易造成果實畸形并伴有不同程度風味和品質的下降,影響商品率和種植效益。為此,本研究以吉優蜜29號甜瓜為試驗材料,采用不同人工花粉組合授粉方式,以激素授粉為對照,測定植株與果實的生長狀況、果實內在品質及蔗糖積累關鍵酶活性,分析授粉對植株與果實生長的影響。結果表明:(1)不同授粉方式處理對甜瓜植株的株高、莖粗、生物量、果實質量、硬度產生很大影響,T1處理的甜瓜植株莖粗、生物量、果實質量顯著高于其他處理。CK處理的甜瓜植株株高、果實硬度顯著高于其他處理。(2)不同授粉方式處理對甜瓜果實內在品質各指標間均存在顯著差異,以T1處理效果最佳,其可溶性固形物、可溶性糖、蔗糖、果糖、葡萄糖含量均顯著高于CK處理。(3)T4處理的蔗糖合成酶(SS)活性在果實生長發育過程中相對較高;T1處理的蔗糖磷酸合成酶(SPS)活性在果實生長發育過程中保持較高水平,T1、T2處理的果實酸性轉化酶活性在授粉后40"d達到最高,分別為93.69、88.54"U/g。因此,人工花粉授粉后甜瓜植株的生長狀況與果實品質以T1(花粉活力強×柱頭可授性高)處理組效果最佳。
關鍵詞:授粉;甜瓜;品質;蔗糖合成酶(SS);蔗糖磷酸合成酶(SPS):酸性轉化酶(AI)中圖分類號:S652""""""文獻標志碼:A
Effects"of"Pollination"Methods"on"Fruit"Quality"and"Related"Enzyme"Activities"of"Melon"Jiyoumi"29
ZHANG"Huiming1,2,"ZHANG"Hanying1,2,"LI"Hongli3,"WANG"Shaochun4,"YANG"Fusun1*
1."School"of"Tropical"Agriculture"and"Forestry,"Hainan"University,"Haikou,"Hainan"570228,"China;"2."Hainan"Changjiang"Melon"Technology"Small"Court,"Changjiang,"Hainan"572732,"China;"3."Tropical"Crop"Genetic"Resources"Institute,"Chinese"Academy"of"Tropical"Agricultural"Sciences,"Haikou,"Hainan"571101,"China;"4."Hainan"Wangpin"Agricultural"Technology"Development"Co.,"Ltd.,"Haikou,"Hainan"570000,"China
Abstract:"Melon"is"an"important"cash"crop"in"Hainan,"and"its"pollination"is"mainly"hormone"pollination,"which"is"easy"to"cause"fruit"deformity"and"accompanied"by"different"degrees"of"flavor"and"quality"decline,"affecting"commodity"rate"and"planting"efficiency."Melon"Jiyoumi"29"was"used"as"the"experimental"material."Different"artificial"pollen"combinations"were"used"to"pollinate"to"compare"with"hormone"pollination."The"growth"status"of"plants"and"fruits,"the"internal"quality"of"fruits"and"the"activities"of"key"enzymes"for"sucrose"accumulation,"and"analyzed"the"effects"of"pollination"on"the"growth"of"plants"and"fruits"were"studied."Different"pollination"methods"had"great"effects"on"plant"height,"stem"diameter"and"biomass,"fruit"weight"and"hardness,"and"the"stem"diameter"and"biomass"and"fruit"weight"treated"with"T1"were"significantly"higher"than"those"of"the"other"treatments."The"plant"height"and"fruit"hardness"treated"with"CK"were"significantly"higher"than"those"of"the"other"treatments."There"were"significant"differences"among"different"pollination"methods"on"each"index"of"melon"fruit"internal"quality,"and"T1"treatment"had"the"best"effect,"and"its"soluble"solids,"soluble"sugar,"sucrose"content,"fructose"content"and"glucose"content"were"significantly"higher"than"CK"treatment."The"activity"of"sucrose"synthase"(SS)"treated"with"T4"was"relatively"high"during"fruit"growth"and"development."The"sucrose"phosphate"synthase"(SPS)"in"T1"treatment"maintained"a"high"level"during"the"fruit"growth"and"development,"and"the"acid"invertase"activity"in"T1"and"T2"treatment"reached"the"highest"at"the"40th"day"after"pollination,"which"were"93.69"U/g"and"88.54"U/g"respectively."The"treatment"of"T1"(strong"pollen"vitality"×"high"stigma"receptivity)"has"the"best"effect"on"the"growth"and"fruit"quality"of"melon"plants"after"artificial"pollen"pollination.
Keywords:"pollination;"Cucumis"melo;"quality;"sucrose"synthase"(SS);"sucrose"phosphate"synthase"(SPS):"acid"invertase"(AI)
DOI:"10.3969/j.issn.1000-2561.2025.05.016
甜瓜(Cucumis"melo"L.)起源于非洲[1],屬葫蘆科(Cucurbitaceae)甜瓜屬(Cucumis)一年生蔓生草本植物[2],其種植區域廣泛,遍布五大洲[3],是一種高經濟的水果作物[4]。目前,甜瓜已成為海南省的主要經濟作物,海南部分地方鄉村振興的重要作物,一年可種植和收獲2~3次[5]。
在植物生長和果實發育的過程中,授粉不僅是種子形成的必要條件,同時也對果實的品質有著重要影響。授粉過程是農作物進行有性繁殖的開始,也是農業生產上最關鍵的一環[6]。當前生產上甜瓜主要依靠激素授粉和自然蜜蜂授粉。激素授粉主要采用氯吡脲(CPPU)進行處理,常因激素劑量和使用時期不當而造成果實畸形,同時影響果實口感與風味[7],并伴隨著不同程度的農藥殘留[8];蜜蜂授粉是早期甜瓜種植的主要傳粉方式,其省時省工、且利于產品安全生產[9],海南等熱區甜瓜種植于大棚,溫度高,蜜蜂授粉易造成蜂群的大量死亡和甜瓜植株早衰,影響產量。有研究表明,人工花粉授粉雖然提高了人工成本,但利于提高果實品質和經濟價值[10-12]。
授粉作為植物生殖生長的關鍵環節,對果實的發育及糖分積累具有重要影響。研究表明,果實風味的形成與果實糖含量及各糖分所占比重關系密切,甜瓜果實中糖積累的主要形式為葡萄糖、果糖和蔗糖[13],而蔗糖的積累是甜瓜果實糖分提高的重要原因[14]。在大多數植物中,蔗糖是源器官(葉)進行光合作用的主要產物,蔗糖合成后會通過韌皮部長距離運輸轉運至果實、根、莖尖等不同的庫器官中維持其生長發育[15-16]。果實中蔗糖代謝的相關酶包括蔗糖合成酶(SS)、蔗糖磷酸化酶(SPS)、酸性轉化酶(AI)[17],蔗糖合成酶可以通過催化葡萄糖和果糖的合成來促進蔗糖的合成,又可以催化蔗糖分解[18-19]。酸性轉化酶促進蔗糖不可逆的分解,蔗糖磷酸合成酶可以將果糖和葡萄糖轉化為蔗糖而在植物體內積累[20]。
蔗糖是植物體內重要的能源和碳源物質,對于植物的生長和代謝具有重要作用。轉化酶參與葉片光合作用,細胞對環境的響應及植物生長發育的調節[21-22],在光合組織中,蔗糖的合成與分配對于維持光合作用的平衡和調節植物體內碳同化的平衡也具有重要意義,這些酶在調控果實內糖分的合成、轉運與代謝中發揮著至關重要的作用[23]。授粉作為果實發育的觸發器,通過影響激素平衡、營養物質分配及基因表達模式,間接或直接調節這些關鍵酶的活性,進而影響甜瓜的糖分積累。而激素與蜜蜂授粉是如何影響甜瓜品質及糖分積累的差異?為此,本研究通過采用人工花粉授粉來模擬蜜蜂授粉方式,對比激素授粉的吉優蜜29號甜瓜,分析不同授粉處理下果實發育過程中糖代謝相關酶活性的動態變化,并結合表型指標及內在品質,探討授粉方式如何調控這些酶的活性,進而影響甜瓜蔗糖代謝。通過調控授粉方式優化甜瓜栽培管理、為提升果實品質提供科學依據,對甜瓜產業的可持續發展具有重要意義。
1""材料與方法
1.1""材料
試驗地點位于海南省昌江黎族自治縣海尾鎮五大村王品果業有限公司蜜瓜試驗示范基地(19.44°N,108.87°E),供試甜瓜品種為吉優蜜29號,試驗時間為2023年12月至2024年5月,每年種植2批,第一批種植時間為同年的8—12月、第二批為12月至次年4月。種植株距36"cm,行距80"cm,種植密度1560株/667"m2。
1.2""方法
1.2.1""試驗設計""試驗采用花粉不同活力與柱頭不同可授性進行人工花粉組合授粉處理(表1)。先測定出吉優蜜29號甜瓜開花規律,確定開花周期。并在開花前1"d進行套袋處理,做好標記。按照表1設置粉源組合,在第15~16個節位的雌花開花時進行人工花粉組合授粉;激素授粉(CK)處理組在開花前1"d進行套袋處理,做好標記。配置好氯吡脲(四川施特優化工有限公司)和咯菌腈(先正達南通作物保護有限公司)混合均勻溶液,在雌花開花時進行激素授粉,將配置好的激素混合溶液用噴壺均勻噴施在雌花瓜胎表面,并進行套袋處理,每個處理3株,3次重復。
1.2.2""指標測定""(1)甜瓜表型、生物量及可溶性固形物測定。在授粉后10、20、30、40"d取樣,采用卷尺、DL91200數顯游標卡尺測定植株株高、莖粗,采用電子天平測定果實質量,使用帶有針形探針(直徑5"mm)的TA"TOUCH"10020質構儀(上海保圣實業發展有限公司)測定硬度,從每個去皮甜瓜陰陽兩面的赤道軸上的等距點獲得2個讀數。植株全生物量測定:收獲時將水沖刷植株根部土壤,取完整植株,105"℃殺青30"min后60"℃烘干至恒重,用百分之一天平準確稱取植株干質量;使用FW-80T糖度計測得甜瓜果實可溶性固形物含量。
(2)葉綠素含量測定。不同授粉天數取樣,測定功能葉的葉綠素,葉綠素含量測定參照95%無水乙醇浸提法[17]進行,暗處放置12~24"h,葉片全部變白后測定葉綠素含量。
(3)果實內在品質測定。不同授粉天數取樣,切塊液氮速凍。將甜瓜果實打磨成粉,保存于50"mL離心管中,置于–80"℃超低溫冰箱待用,按照酶試劑盒說明書測定甜瓜果實的可溶性糖、果糖、葡萄糖和蔗糖含量,3次重復,平行3次,測定均使用北京索萊寶科技有限公司試劑盒。
(4)糖分轉化相關酶活性測定。不同授粉天數取樣,切塊液氮速凍。將甜瓜果實打磨成粉,保存于50"mL離心管中,置于–80"℃超低溫冰箱待用。按照酶試劑盒說明書測定甜瓜果實的蔗糖合成酶(SS)、蔗糖磷酸合成酶(SPS)和酸性轉化酶(AI)活性,3次重復,平行3次,測定均使用北京索萊寶科技有限公司試劑盒。
1.3""數據處理
采用Excel"2010軟件整理數據,采用SPSS"Statistics"26軟件統計分析各成分的差異性,并進行單因素方差分析(one-way"ANO-VA),采用Origin"2021軟件繪圖。
2""結果與分析
2.1""授粉方式對甜瓜生長和生物量的影響
不同授粉方式授粉40"d后,植株株高、莖粗、果實質量、果實硬度、生物量間均差異顯著(表2)。人工花粉4個處理組間,T1處理的甜瓜植株莖粗、果實質量、生物量顯著高于T2、T3、T4處理,T2、T3、T4處理間差異不顯著;CK處理的植株株高高于T1和T4處理,且差異顯著,果實硬度顯著高于其他處理。
2.2""授粉方式對甜瓜功能葉光合色素含量的影響
不同授粉方式處理后,隨著授粉天數的增加,甜瓜功能葉葉綠素b含量呈先上升后下降的趨勢(圖1)。CK處理下葉綠素b含量在20"d時達到最大,且與其他處理差異顯著,較T1處理增加2.93%,較T2處理增加24.55%,較T3處理增加8.44%,較T4處理增加18.12%;T2處理下葉綠素b含量在20~40"d降幅最小,較T1處理減少31.69%,較T3處理減少29.33%,較T4處理減少17.04%,較CK處理減少18.46%。
2.3""授粉方式對甜瓜生理影響的相關性分析
由圖2可知,不同授粉方式處理對甜瓜植株
和果實表型性狀、光合色素含量間相關性較高,植株株高與莖粗、單果質量、生物量呈顯著負相關,相關系數分別為–0.55、–0.58、–0.63,與葉綠素b含量呈顯著正相關,相關系數為0.53;單果質量與生物量呈顯著正相關,相關系數為0.91;葉綠素a含量與葉綠素b含量呈顯著正相關,相關系數為0.83。
2.4""授粉方式對甜瓜果實內在品質的影響
不同授粉方式授粉40"d后,果實內在品質各指標間均差異顯著(表3)。人工花粉4個處理組間,以T1處理效果最佳,可溶性固形物、蔗糖含量和果糖含量顯著高于其他處理;不同授粉方式處理的果實葡萄糖含量差異不顯著。
2.5""授粉方式對甜瓜糖分積累相關酶活性影響
2.5.1""蔗糖合成酶(SS)活性""不同授粉方式處理下果實中SS活性總體呈現出“升-降-升”的變化趨勢(圖3)。授粉10、40"d時,CK處理的SS活性顯著高于其他處理;授粉20"d,T4處理下SS活性高達576.57"U/g,顯著高于其他處理;授粉30"d,T1和T2處理下SS活性達到最低,顯著低于其他處理。
2.5.2""蔗糖磷酸合成酶(SPS)活性""由圖4可知,不同授粉方式處理下果實中SPS活性總體呈現出“降-降-升”的變化趨勢,而T3處理下甜瓜果實SPS活性呈“升-降-升”的趨勢,并在授粉20"d時達到最高,較其他處理增幅為29.51%~32.23%,在授粉30"d時達到最低,顯著低于其他處理。
2.5.3""酸性轉化酶(AI)活性""不同授粉方式處理下甜瓜果實中SPS活性總體呈現先下降后上升的變化趨勢,各處理在不同授粉天數間均差異顯著(圖5)。T1、T2處理組在授粉40"d時AI活性達到最高,分別為93.69、88.54"U/g,且與其他處理存在顯著性差異;CK處理在授粉10、40"d時AI活性均低于其他處理,分別為52.83、21.34"U/g,且差異顯著。
2.5.4""甜瓜果實內在品質與蔗糖代謝關鍵酶活性相關性""由圖6可知,不同授粉方式處理對甜瓜果實內在品質與蔗糖代謝關鍵酶含量間相關性較高,可溶性固形物與蔗糖含量、酸性轉化酶呈顯著正相關,相關系數分別為0.60、0.74,與蔗糖合成酶呈顯著負相關;蔗糖含量與蔗糖合成酶呈顯著負相關,相關系數為–0.69;果糖含量與蔗糖磷酸合成酶呈顯著正相關,相關系數為0.79;蔗糖合成酶與酸性轉化酶呈顯著負相關,相關系數為–0.84。
3""討論
花粉活力和授粉時機的把握對于植物生長發育具有重要意義?;ǚ刍盍椭^可授性的強弱是決定授粉成功率的關鍵因素[24],它不僅影響授粉后花粉管的伸長和受精過程,還與果實的品質和產量密切相關。不同授粉方式授粉40"d,T1(花粉活力強×柱頭可授性高)處理的甜瓜植株與果實長勢良好,優于其他處理,而CK(激素授粉)的植株株高和果實硬度顯著高于人工花粉授粉處理組,激素的合理使用可以有效地促進植物的生長發育。同時,生物量是反映植物體有機物積累狀況的主要指標[25],是作物產量和品質形成的基礎[26]。本研究中,T1處理的植株生物量顯著高于其他處理,說明T1處理的果實具有較高的生長速度、積累的有機物較多,使得積累生物量顯著高于其他處理。
葉綠素作為植物進行光合作用的主要色素,其含量通常與光合作用呈正相關[27],光合作用是植物進行生產的基礎[28]。不同授粉方式處理下,甜瓜功能葉葉綠素含量20"d時達到最高值,之后開始降低,可能是由于甜瓜果實在生長發育過程中,隨著果實的膨大,功能葉葉片也在逐漸增大,使得葉綠素含量在0~20"d時達到最高,隨著果實的發育成熟,功能葉葉片停止生長轉而逐漸衰老,直至枯萎,葉綠素含量在40"d時降到最低。
在人工花粉授粉過程中,花粉的活力和柱頭的可授性是影響果實品質的關鍵因素。高活力花粉和良好可授性的柱頭有助于果實營養成分的顯著提升。其中,可溶性固形物和可溶性糖是衡量果實口感和營養價值的重要指標,可溶性固形物的增加將帶來更佳的口感和更豐富的營養,可溶性糖是調控植物生長發育的重要信號分子[20]。本研究表明,人工花粉授粉的甜瓜其可溶性固形物和可溶性糖含量顯著高于激素授粉。
蔗糖合成酶(SS)和蔗糖磷酸合成酶(SPS)在蔗糖的合成過程中起著關鍵作用[27]。SS主要催化蔗糖的合成,將單糖轉化為蔗糖,而SPS則通過磷酸化作用促進蔗糖的合成[29]。這2種酶在植物體內相互協作,共同調節蔗糖的合成速率和方向,酸性轉化酶則主要參與蔗糖的分解過程。在植物體內,酸性轉化酶可以將蔗糖分解為單糖,從而影響蔗糖的運輸和利用[30]。此外,它還可以將光合作用合成的蔗糖進行分配,用于儲存、呼吸或其他生物合成過程[31]。本研究中,不同授粉方式處理對甜瓜糖分積累相關酶活性有不同程度的影響,T4處理的蔗糖合成酶活性在果實生長發育過程中相對較高,T1處理的蔗糖磷酸合成酶在果實生長發育過程中保持較高水平,T1、T2處理的果實酸性轉化酶活性在授粉后40"d時達到最高,可能是由于T1、T2處理的甜瓜成熟期早,采摘后快速進入后熟階段,使得酸性轉化酶快速增加,而酸性轉化酶的增加有助于果實的成熟和軟化[32],通過分解蔗糖,釋放出的葡萄糖和果糖可以為果實的細胞提供能量,促進細胞壁的降解和果肉的軟化,從而使果實更易于食用和消化[33-34]。
4""結論
不同授粉方式對甜瓜植株的生長形態、生理過程存在明顯的相關性,即T1處理組在授粉后,甜瓜植株的生長性狀和光合色素含量在10~30"d時保持較高水平,顯著高于其他處理,長勢最佳,隨著果實的成熟,在40"d時達到最低,果實成熟期早;同時不同授粉方式對甜瓜果實品質及糖分積累關鍵酶含量間相關性較高,T1處理的甜瓜果實內在品質顯著高于其他處理。人工花粉授粉4個處理組的蔗糖合成酶、蔗糖磷酸合成酶、酸性轉化酶,在整個果實生長發育過程中均保持在較高水平;不同授粉方式授粉后,甜瓜果實的蔗糖含量與蔗糖磷酸合成酶、酸性轉化酶呈顯著正相關、與蔗糖合成酶呈顯著負相關;蔗糖合成酶與酸性轉化酶呈顯著負相關,綜合考慮甜瓜的生長特性、內在品質及糖分積累相關酶的活性,得出甜瓜在T1(花粉活力強×柱頭可授性高)授粉后生長狀況與果實品質最佳。
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