周睿琦,談倩倩,王小平,劉 文
(華中農(nóng)業(yè)大學(xué)植物科學(xué)技術(shù)學(xué)院,昆蟲資源利用與害蟲可持續(xù)治理湖北省重點(diǎn)實(shí)驗(yàn)室,武漢 430070)
大猿葉蟲滯育品系和非滯育品系交配行為的比較
周睿琦,談倩倩,王小平,劉 文*
(華中農(nóng)業(yè)大學(xué)植物科學(xué)技術(shù)學(xué)院,昆蟲資源利用與害蟲可持續(xù)治理湖北省重點(diǎn)實(shí)驗(yàn)室,武漢 430070)
大猿葉蟲ColaphellusbowringiBlay是一種重要的十字花科蔬菜害蟲,以成蟲越冬和越夏。明確大猿葉蟲非滯育品系生殖力下降的現(xiàn)象是否與交配行為的改變相關(guān),將促進(jìn)該非滯育品系在滯育遺傳、滯育調(diào)控分子機(jī)制研究中的應(yīng)用。本研究在25℃和L ∶D=12 ∶12 h條件下,測(cè)定了大猿葉蟲滯育品系和非滯育品系的交配節(jié)律、交配持續(xù)時(shí)間和交配頻次。結(jié)果表明,大猿葉蟲滯育品系和非滯育品系交配高峰均出現(xiàn)在光期開始后2 h,非滯育品系的交配比率顯著低于滯育品系。滯育品系和非滯育品系第1次交配持續(xù)時(shí)間無顯著差異,分別為122.7±7.4 min和100.1±10.2 min;但非滯育品系的總交配持續(xù)時(shí)間顯著低于滯育品系。非滯育品系的交配頻次為2.2 ±0.3次,顯著低于滯育品系的3.8±0.5次。滯育品系和非滯育品系交配行為的比較發(fā)現(xiàn),大猿葉蟲非滯育品系生殖力降低與其交配比率下降、交配持續(xù)時(shí)間縮短和交配頻次減少顯著相關(guān),本研究為進(jìn)一步利用非滯育品系為材料解析滯育遺傳、滯育調(diào)控機(jī)制奠定了基礎(chǔ)。
大猿葉蟲;非滯育品系;交配節(jié)律;交配持續(xù)時(shí)間;交配頻次
滯育(Diapause)是昆蟲應(yīng)對(duì)外界不良環(huán)境的一種季節(jié)性適應(yīng)策略,主要表現(xiàn)為營養(yǎng)代謝模式的轉(zhuǎn)變和發(fā)育停滯(Tauberetal.,1986)。滯育昆蟲具有遺傳多樣性,其滯育能力(diapause capacity)可被自然或人工選擇改變。通過自然選擇或人工篩選已在40多種昆蟲和螨類中獲得非滯育品系(Tauberetal.,1986;Riihimaa and Kimura,1988;Ito,2009)。有的昆蟲非滯育品系已被用于滯育遺傳研究的材料,例如,比較始紅蝽Pyrrhocorisapterus滯育品系和非滯育品系雜交、回交后親代和子代的光周期反應(yīng),最終探明了始紅蝽的滯育綜合征(diapause syndrome)是顯性性狀(Socha and Hodková,1994)。有的昆蟲非滯育品系可被用于害蟲遺傳防治,田間釋放害蟲的非滯育品系,非滯育品系個(gè)體與野生品系個(gè)體雜交后,可改變害蟲的基因型,在不利環(huán)境到來時(shí),害蟲無法進(jìn)入滯育而死亡(Hoy and Knop,1978)。然而,很多昆蟲非滯育品系存在適應(yīng)性退化的現(xiàn)象,其中生殖力降低最為明顯(Tauberetal.,1986)。例如,在守瓜Atrachyamenetriesi中,非滯育品系雌成蟲的產(chǎn)卵量下降,卵的孵化率降低(Ando and Miya,1968)。交配是兩性生殖昆蟲繁衍后代的重要環(huán)節(jié),交配節(jié)律、交配持續(xù)時(shí)間和交配頻次均可能影響昆蟲的生殖力(Arnqvist and Nilsson,2000;Herbersteinetal.,2010)。有研究表明,昆蟲非滯育品系的生殖力降低可能與交配行為的改變有關(guān)(Herbersteinetal.,2010),但非滯育品系的哪些交配行為發(fā)生改變進(jìn)而影響到其生殖力尚不清楚。
大猿葉蟲ColaphellusbowringiBaly是一種重要的十字花科蔬菜害蟲,低溫或長光照會(huì)誘導(dǎo)其成蟲入土滯育(薛芳森等,2002;Xueetal.,2002)。前期報(bào)道過一個(gè)對(duì)光周期不敏感的大猿葉蟲非滯育品系,該非滯育品系在溫度高于20℃時(shí),無論何種光周期均不能誘導(dǎo)個(gè)體進(jìn)入滯育(Maetal.,2011;Tanetal.,2016)。大猿葉蟲非滯育品系是研究滯育遺傳的良好材料,利用大猿葉蟲滯育品系與非滯育品系雜交,已探明光周期誘導(dǎo)的滯育是不完全顯性遺傳且受母本效應(yīng)影響較大(Kuangetal.,2011;Maetal.,2011)。然而,大猿葉蟲非滯育品系的生殖力明顯降低,其雌成蟲壽命、產(chǎn)卵量和卵孵化率均顯著低于正常品系(Tanetal.,2016)。前期觀察發(fā)現(xiàn),大猿葉蟲交配行為中的交配頻次和交配持續(xù)時(shí)間顯著影響著其生殖力,交配頻次與其產(chǎn)卵量正相關(guān),較長的交配持續(xù)時(shí)間可提高產(chǎn)卵量(Liuetal.,2010;Liuetal.,2013)。因此,推測(cè)大猿葉蟲非滯育品系生殖力的降低可能與其交配行為的改變有關(guān),但非滯育品系的交配行為與滯育品系相比存在哪些差異仍不清楚。
為明確大猿葉蟲非滯育品系生殖力降低的原因,在25℃和L ∶D=12 ∶12 h條件下,分別測(cè)定大猿葉蟲滯育品系和非滯育品系的交配節(jié)律、交配持續(xù)時(shí)間和交配頻次。研究結(jié)果可明確大猿葉蟲非滯育品系生殖力降低與其交配行為發(fā)生改變的相關(guān)性,為以非滯育品系為材料研究昆蟲滯育遺傳、滯育調(diào)控機(jī)制提供科學(xué)依據(jù)。
1.1 供試?yán)ハx
2008年11月在江西省修水縣(29°1′N,114°4′E)采集超過1000頭的大猿葉蟲成蟲,在自然條件下的玻璃缸中入土滯育(Xueetal.,2002)。大猿葉蟲成蟲滯育解除后子代有明顯的光周期反應(yīng),溫度高于20℃時(shí)長光照誘導(dǎo)其進(jìn)入滯育,視為滯育品系(high-diapause strain,HD strain)。大猿葉蟲滯育解除后,將成蟲轉(zhuǎn)移到塑料養(yǎng)蟲盒(16 cm × 7.5 cm × 6 cm)內(nèi)繁殖,用于試驗(yàn)。
2006年11月在江西省修水縣(29°1′N,114°4′E)采集500-600頭大猿葉蟲成蟲,將滯育解除后的成蟲飼養(yǎng)在非滯育誘導(dǎo)條件下(22℃和L ∶D=13 ∶11),部分成蟲未進(jìn)入滯育。將未滯育成蟲飼養(yǎng)在滯育誘導(dǎo)條件22℃和L ∶D=16 ∶8下,持續(xù)飼養(yǎng)篩選22代后,滯育比率低于5%,對(duì)光周期誘導(dǎo)失去敏感性,獲得了較穩(wěn)定的非滯育品系(non-diapause strain,NDS)(Maetal.,2011),該非滯育品系在室內(nèi)22℃和L ∶D=16 ∶8 h條件下連續(xù)飼養(yǎng)保存。
1.2 試驗(yàn)條件
所有幼蟲或成蟲放入塑料養(yǎng)蟲盒內(nèi),以新鮮蘿卜Raphanusstativusvar.longipinnatus葉飼養(yǎng)。飼養(yǎng)條件為溫度25℃±1℃、光周期L ∶D=12 ∶12 h、相對(duì)濕度50%-60%、光照強(qiáng)度500-700 Lx(薛芳森等,2002)。飼養(yǎng)的全過程均在光照培養(yǎng)箱(HP250GS,武漢瑞華儀器設(shè)備有限責(zé)任公司)中進(jìn)行,暗期處理采用人工套袋(在養(yǎng)蟲盒外加套不透光的黑布袋,至光期再移除)的方法進(jìn)行(Xueetal.,2002)。
1.3 交配行為觀察
將單頭滯育品系或非滯育品系5日齡未交配雌成蟲放入墊有保濕濾紙和新鮮蘿卜葉的培養(yǎng)皿(直徑 5.0 cm)中,適應(yīng)10 min后,引入1頭同品系的5日齡未交配雄成蟲。大猿葉蟲交配多發(fā)生在光期,雌雄成蟲可多次交配,且第1次交配持續(xù)時(shí)間顯著長于之后的交配持續(xù)時(shí)間(Liuetal.,2010)。配對(duì)后立即觀察成蟲的交配情況,觀察完整的光期,從9 ∶00開始至21 ∶00結(jié)束,連續(xù)觀察12 h。每對(duì)雌雄成蟲為1次重復(fù),滯育品系和非滯育品系觀察樣本分別為30對(duì)和40對(duì)雌雄成蟲。記錄每對(duì)雌雄成蟲的交配起始時(shí)間、交配結(jié)束時(shí)間、交配頻次(Liuetal.,2010),同一對(duì)雌雄成蟲的總交配持續(xù)時(shí)間為其多次交配持續(xù)時(shí)間之和。
1.4 數(shù)據(jù)分析
試驗(yàn)中的數(shù)據(jù)均采用SPSS 11.5進(jìn)行分析。對(duì)大猿葉蟲滯育品系和非滯育品系雌雄成蟲觀察對(duì)數(shù)和交配對(duì)數(shù)進(jìn)行卡方檢驗(yàn),比較二者間交配比率的差異;交配持續(xù)時(shí)間、交配頻次的平均值差異通過t測(cè)驗(yàn)(t-test,P<0.05)進(jìn)行分析。
圖 1 大猿葉蟲滯育品系和非滯育品系的交配節(jié)律Fig.1 The mating rhythm of the diapause strain (DS)and non-diapause strain (NDS)of Colaphellus bowringi
2.1 交配節(jié)律
在25℃和L ∶D=12 ∶12 h條件下,大猿葉蟲滯育品系和非滯育品系的交配高峰均出現(xiàn)在光期開始后2 h,交配率分別為76.7%和30.0%。滯育品系的交配率為90.0%,非滯育品系為47.5%,大猿葉蟲滯育品系的交配率顯著高于非滯育品系(χ2=42.128,P<0.01)(圖1)。
2.2 交配持續(xù)時(shí)間
圖2 大猿葉蟲滯育品系和非滯育品系交配持續(xù)時(shí)間的分布Fig. 2 The distribution of the mating duration of diapause strain (DS)and non-diapause strain (NDS)of Colaphellus bowringiA,第一次交配持續(xù)時(shí)間;B,總交配持續(xù)時(shí)間。A, mating duration for the first time; B, total mating duration.
大猿葉蟲滯育品系和非滯育品系第1次交配持續(xù)時(shí)間的分布相似,超過60%的個(gè)體第1次交配持續(xù)時(shí)間大于90 min(圖2A)。大猿葉蟲滯育品系和非滯育品系總交配持續(xù)時(shí)間分布存在明顯差異,滯育品系總交配持續(xù)時(shí)間最短為73 min,最長達(dá)到417 min;而非滯育品系總交配持續(xù)時(shí)間集中在90-150 min(圖2B)。滯育品系第1次交配持續(xù)時(shí)間為122.7±7.4 min,非滯育品系的第1次交配持續(xù)時(shí)間為100.1±10.2 min,兩品系第1次交配持續(xù)時(shí)間無顯著性差異(t=1.847,P=0.072)(圖3A);大猿葉蟲滯育品系總交配持續(xù)時(shí)間為232.6±18.7 min,非滯育品系總交配持續(xù)時(shí)間為152.3±20.1 min,滯育品系總交配持續(xù)時(shí)間顯著長于非滯育品系(t=2.877,P<0.01)(圖3B)。
圖3 大猿葉蟲滯育品系和非滯育品系交配持續(xù)時(shí)間的比較Fig.3 The comparison of mating duration between diapause strain (DS)and non-diapause strain (NDS)of Colaphellus bowringi注:A,第一次交配持續(xù)時(shí)間;B,總交配持續(xù)時(shí)間。Note:A, mating duration for the first time; B, total mating duration.
2.3 交配頻次
大猿葉蟲滯育品系和非滯育品系交配頻次的分布存在明顯差異,大猿葉蟲滯育品系大部分個(gè)體1天中交配頻次大于3次,而非滯育品系大部分個(gè)體1天中交配1-2次。大猿葉蟲滯育品系總交配頻次為3.8±0.5次,而非滯育品系總交配頻次為2.2±0.3次,滯育品系交配頻次顯著多于非滯育品系(t=2.481,P< 0.001)(圖4)。
圖4 大猿葉蟲滯育品系和非滯育品系平均交配頻次Fig.4 Meanfrequency of mating between diapause strain (DS) and non-diapause strain (NDS)of Colaphellus bowringi
昆蟲非滯育品系在滯育遺傳研究和害蟲生物防治中有潛在應(yīng)用價(jià)值,但大部分昆蟲非滯育品系存在生殖力降低的現(xiàn)象(Tauberetal.,1986)。本研究對(duì)大猿葉蟲滯育品系和非滯育品系交配行為的比較發(fā)現(xiàn),滯育品系和非滯育品系的交配節(jié)律無明顯差異,但非滯育品系的交配率顯著下降、總交配持續(xù)時(shí)間顯著縮短,交配頻次顯著減少,說明非滯育品系的交配能力較之滯育品系顯著降低,這為明確大猿葉蟲非滯育品系生殖力降低的內(nèi)在機(jī)制奠定了基礎(chǔ)。
大猿葉蟲交配多發(fā)生在光期,雌雄成蟲可多次交配,第1次交配持續(xù)時(shí)間顯著長于之后的交配持續(xù)時(shí)間(Liuetal.,2010)。本研究表明,大猿葉蟲滯育品系和非滯育品系第1次交配持續(xù)時(shí)間無顯著差異,但非滯育品系的總交配持續(xù)時(shí)間顯著短于滯育品系。在昆蟲中,交配持續(xù)時(shí)間的長短對(duì)其生殖適應(yīng)性有重要影響(Herbersteinetal.,2010)。在多數(shù)昆蟲中,雄成蟲通過延長交配持續(xù)時(shí)間來增加可向雌成蟲轉(zhuǎn)移的精子數(shù),同時(shí)延長交配持續(xù)時(shí)間可作為一種配偶保護(hù)行為,避免精子競(jìng)爭(zhēng)(Thornhill and Alcock,1983;Simmons,2001);對(duì)于雌成蟲,延長交配持續(xù)時(shí)間有利于貯存更多的精子和隨精子一同轉(zhuǎn)移的營養(yǎng)物質(zhì)進(jìn)而提高生殖成功的幾率(Arnqvist and Nilsson,2000)。交配持續(xù)時(shí)間對(duì)產(chǎn)卵量的影響在許多昆蟲中已有報(bào)道。例如對(duì)于茄28星瓢蟲Henosepilachnavigintloctopunctata,較長的交配持續(xù)時(shí)間導(dǎo)致更高的產(chǎn)卵量和卵孵化率(周康念等,2012)。因此,大猿葉蟲非滯育品系總交配持續(xù)時(shí)間的縮短,可能導(dǎo)致雌成蟲從雄成蟲精液獲得的營養(yǎng)物質(zhì)和精子數(shù)減少,不利于雌成蟲的生殖,最終導(dǎo)致其非滯育品系雌成蟲產(chǎn)卵量的下降。
許多昆蟲在一個(gè)生殖季節(jié)中能發(fā)生多次交配(multiple mating),這種多次交配常有利于提高雌成蟲的生殖適應(yīng)性(Arnqvist and Nilsson,2000)。雌成蟲通過多次交配能獲得更多的營養(yǎng)進(jìn)而刺激生殖行為,提高生殖產(chǎn)量;同時(shí),多次交配也有利于雌成蟲儲(chǔ)存足夠的精子,確保所有卵的受精(Thornhill and Alcock,1983;Arnqvist and Nilsson,2000;孟玲和李保平,2006)。在大猿葉蟲中,雌成蟲交配頻次與其產(chǎn)卵量呈正相關(guān)(Liuetal.,2013)。大猿葉蟲非滯育品系1天內(nèi)交配頻次顯著少于滯育品系,交配頻次的減少與其總交配持續(xù)時(shí)間的縮短有直接關(guān)系。因此,交配頻次減少可能是導(dǎo)致大猿葉蟲非滯育品系生殖力降低的關(guān)鍵因素。
本研究表明大猿葉蟲非滯育品系較之滯育品系,具有低的交配率、短的交配持續(xù)時(shí)間、少的交配頻次,這些均有可能導(dǎo)致非滯育品系雌成蟲的生殖力下降,從而導(dǎo)致非滯育品系適應(yīng)性降低。明確大猿葉蟲非滯育品系生殖力降低的原因與其交配行為發(fā)生改變的相關(guān)性,為進(jìn)一步利用大猿葉蟲非滯育品系為材料分析昆蟲滯育遺傳、滯育調(diào)控的機(jī)制提供了依據(jù)。
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Differences inmating behavior between diapause and non-diapause strains of the cabbage beetleColaphellusbowringi(Coleoptera: Chrysomelidae)
ZHOU Rui-Qi, TAN Qian-Qian, WANG Xiao-Ping, LIU Wen*
(Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China)
The cabbage beetleColaphellusbowringiBaly (Coleoptera: Chrysomelidae)is a serious pest of crucifers and goes into summer or winter diapause as adults. Compared to the diapause strain (DS), the reduced fecundity of non-diapause strain (NDS)is possibly related to mating behavior. The NDS ofC.bowringiis a promising model for investigating the mechanism of diapause inheritance and regulation. In this study, we observed the mating rhythm, mating duration and mating frequency of the DS and NDS inC.bowringiat 25℃ and L ∶D=12 ∶12 h. The results showed that mating rhythm of both strains peaked at 2 h after light period began, and the mating rate of NDS was significant lower than that of DS. The mating durations of the first time of DS and NDS were 122.7±7.4 min and 100.1±10.2 min, respectively, and there is no significant difference. However, the total mating duration of NDS was significant lower than that of DS. The mating frequency of DS was 3.8±0.5 times which was significant higher than that of NDS (2.2 ± 0.3 times). Comparison of the mating behavior between DS and NDS shows that the reduced fecundity of NDS is associated with the mating behavior variation, including decreased mating rate, shortened mating duration and reduced mating frequency. The application of NDS can promote the study of the mechanism of insect diapause inheritance and regulation.
Colaphellusbowringi; non-photoperiodic-diapause strain; mating rhythm; mating duration; mating frequency
國家自然科學(xué)基金(31272045, 31572009);中央高?;究蒲袠I(yè)務(wù)費(fèi)專項(xiàng)資金(2014BQ012)
周睿琦,女,1991年生,湖北武漢人,農(nóng)學(xué)碩士,研究方向?yàn)槔ハx生態(tài)學(xué),E-mail:richchou19911017@163.com
*通訊作者 Author for correspondence,E-mail: liuwen@mail.hzau.edu.cn
Received:2016-08-16; 接受日期Accepted: 2016-10-22
Q968.1;S433
A
1674-0858(2017)01-0167-05
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