团头鲂(♀)×翘嘴鲌(♂)杂交三倍体的生长性能及形态分析

引用本文

张亮华, 贾橙钰, 许文雅, 徐文婧, 郑国栋, 邹曙明. 团头鲂(♀)×翘嘴鲌(♂)杂交三倍体的生长性能及形态分析[J]. 渔业科学进展, 2024, 45(5): 134-143. DOI: 10.19663/j.issn2095-9869.20230612001.

ZHANG Lianghua, JIA Chengyu, XU Wenya, XU Wenjing, ZHENG Guodong, ZOU Shuming. Growth Performance and Morphological Analysis of Triploids of Megalobrama amblycephala (♀) × Culter alburnus (♂)[J]. Progress in Fishery Sciences, 2024, 45(5): 134-143. DOI: 10.19663/j.issn2095-9869.20230612001.

基金项目

上海市科技兴农重点攻关项目((2022-02-08-00-12-F01179)、上海高校青年教师培养资助计划重点项目(A1-2007-23-000203)、国家重点研发计划(2022YFD2400602)和上海海洋大学青年教师科研启动经费(A2-2006-22-200305)共同资助

作者简介

张亮华，Email: 1312566966@qq.com

通讯作者

邹曙明，教授，Email: smzou@shou.edu.cn

文章历史

收稿日期：2023-06-12
收修改稿日期：2023-07-01

Contents Abstract Full text Figures/Tables PDF

团头鲂(♀)×翘嘴鲌(♂)杂交三倍体的生长性能及形态分析

张亮华 , 贾橙钰 , 许文雅 , 徐文婧 , 郑国栋 , 邹曙明

上海海洋大学农业农村部团头鲂遗传育种中心农业农村部淡水水产种质资源重点实验室水产科学国家级实验教学示范中心上海 201306

收稿日期：2023-06-12；收修改稿日期：2023-07-01

基金项目：上海市科技兴农重点攻关项目((2022-02-08-00-12-F01179)、上海高校青年教师培养资助计划重点项目(A1-2007-23-000203)、国家重点研发计划(2022YFD2400602)和上海海洋大学青年教师科研启动经费(A2-2006-22-200305)共同资助

作者简介：张亮华，Email: 1312566966@qq.com.

通讯作者：邹曙明，教授，Email: smzou@shou.edu.cn.

摘要：鲂鲌(Megalobrama amblycephala♀ × Culter alburnus ♂)三倍体因性腺不发育或败育而将更多能量用于生长，因为不育可以保持F₁代的杂种优势而避免继续繁殖造成的种质衰退，同时具备生态安全性，因此，鲂鲌三倍体具有优越的“生长优势”或“经济性状”。为了解鲂鲌三倍体的生长情况与形态特征，本研究运用静水压技术对团头鲂(♀)×翘嘴鲌(♂)杂交受精卵进行三倍体化诱导，获得了诱导成功的鲂鲌三倍体群体(hybrid-3n)与未诱导成功的鲂鲌二倍体群体(hybrid-2n)，并构建团头鲂(MA)和翘嘴鲌(CA) 2个自交群体，通过生长对比实验比较分析了4个群体鱼的生长性能及形态差异。结果显示，在210 d生长周期内，鲂鲌三倍体绝对增重率[(0.88±0.11) g/d]比二倍体提高8.64%，较MA提高20.55%，较CA提高120.00%。可数性状上，鲂鲌三倍体和二倍体杂种指数平均值分别为41.05和36.07，都略偏向于母本团头鲂。可量性状上，鲂鲌三倍体杂种指数平均值为36.73，略偏向于母本团头鲂；鲂鲌二倍体杂种指数平均值为57.57，略偏向于父本翘嘴鲌。聚类分析显示，鲂鲌三倍体首先和二倍体聚在一起归为一类，然后再与母本团头鲂聚为一支，最后再与父本翘嘴鲌聚在一起。判别分析显示，对鲂鲌三倍体的判别准确率为90%，对4个群体的综合判别率达95%。主成分分析共得到累计贡献率达75.10%的4个主成分，主要体现了鱼体高和躯干的形态变异。综合分析表明，鲂鲌三倍体具有较快的生长速度，具备在生产上推广应用的基础条件；鲂鲌三倍体与二倍体体型均介于双亲之间，都略偏向于母本，主要体现了鱼体高和躯干的形态变异。研究结果对鲂鲌杂交后代种群鉴定、倍性育种具有重要意义。

关键词：鲂鲌杂交三倍体生长速度形态特征多元统计

Growth Performance and Morphological Analysis of Triploids of Megalobrama amblycephala (♀) × Culter alburnus (♂)

ZHANG Lianghua , JIA Chengyu , XU Wenya , XU Wenjing , ZHENG Guodong , ZOU Shuming

Genetics and Breeding Center for Blunt Snout Bream, Ministry of Agriculture and Rural Affairs; Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs; National Demonstration Center for Experimental Fisheries Science Eduction; Shanghai Ocean University, Shanghai 201306, China

Corresponding author: ZOU Shuming, Email: smzou@shou.edu.cn.

Abstract: Megalobrama amblycephala, which belongs to the genus Megalobrama of the subfamily Culterinae in the family Cyprinidae, is an important freshwater cultured species in China because of its low culture cost and high growth rate. Culter alburnus, belonging to the subfamily Culter, is another slender carnivorous species, whose meat is more delicate and delicious than that of M. amblycephala; however, the scales of C. alburnus are small and thin and are easily injured and fall off, and these fish are slow-growing and expensive to feed. Although both the fish are quite different in terms of diet, growth, and stress resistance, they are highly complementary to each other. Crossbreeding can combine the desired characteristics of both parents; however, this advantage is usually limited to the F₁ generation. Theoretically, because the gonads are almost undeveloped during triploid growth and development, F₁ heterosis can be maintained to avoid germplasm decline caused by continuous reproduction, and energy can be saved to accelerate growth and improve muscle quality. This will greatly expand the breeding space of new hybrid varieties and ensure high economic, social, and ecological value. Therefore, the triploids of M. amblycephala ♀ × C. alburnus ♂ have superior growth and economic value. To understand the growth and morphological characteristics of the triploids of M. amblycephala ♀ × C. alburnus ♂, the triploid population of M. amblycephala ♀ × C. alburnu ♂ was induced by hydrostatic pressure, and successfully induced triploid (hybrid-3n) and uninduced diploid (hybrid-2n) populations were obtained. At the same time, two self-breeding populations of Megalobrama (MA) and Culter (CA) were established. The growth performance and morphological characteristics of the four kinds of fish were compared and analyzed in growth contrast experiments. The results showed that during the 210-day growth cycle, the absolute weight gain (0.88±0.11 g/day) of hybrid-3n was 8.64% higher than that of hybrid-2n, 20.55% higher than that of MA, and 120.00% higher than that of CA. In terms of countable traits, the hybrid index of hybrid-3n and hybrid-2n was 41.05 and 36.07, respectively. In terms of measurable traits, the hybrid index of hybrid-3n and hybrid-2n was 36.73 and 57.57, respectively. Cluster analysis showed that hybrid-3n was first grouped with hybrid-2n, then grouped with maternal MA, and finally grouped with paternal CA. The results showed that hybrid-3n and hybrid-2n were closer to their mothers in quantifiable traits and frame structure, showing a maternal effect. Discriminant analysis showed that the discriminant accuracy of hybrid-3n was 90%, and the comprehensive discriminant rate of the four populations was 95%. A scatterplot of the discriminant analysis showed that the distribution centers of hybrid-3n and hybrid-2n were located between the parents and closer to the mothers. Hybrid-3n and hybrid-2n occupied partially overlapping areas, which indicates that the two are similar in proportion and frame structure, making it difficult to distinguish between them. These results are basically consistent with those obtained by cluster analysis. Most of the 11 biological traits used to construct the discriminant function were related to the longitudinal axis of the body, especially the ratio of the height of the body to the trunk, which is similar to the results obtained by principal component analysis. During principal component analysis, four principal components with a cumulative contribution rate of 75.10% were obtained, which mainly reflect morphological variations of body height and trunk length. In this study, the experimental fish were cultured in still water ponds with abundant bait. The morphological differences may be attributed to the adaptation of the fish to this ecological environment. Comprehensive analysis showed that: Hybrid-3n has a fast growth rate and has the basic conditions for promotion and application in production; The body size of hybrid-3n and hybrid-2n is between the parents, and both are slightly biased towards the mother, which mainly reflect the morphological variations of body height and trunk length; In this study, three multivariate analyses were used to effectively reflect the morphological differences among the four populations of hybrid-3n, hybrid-2n, MA, and CA from different perspectives, which has made them irreplaceable. In conclusion, this study confirmed the superior breeding potential of triploids of M. amblycephala ♀ × C. alburnu ♂, and provided basic data for the morphological comparison of hybrid offspring of M. amblycephala and C. alburnu, which is significant for the identification and protection of fish resources, and is expected to provide a theoretical basis for the establishment of new strains of M. amblycephala and C. alburnu. The results are of great significance for population identification and ploidy breeding of hybrid offspring of M. amblycephala and C. alburnu.

Key words: Hybrid of Megalobrama amblycephala♀×Culter alburnus♂ Triploid Growth rate Morphological characteristics Multivariate statistics

团头鲂(Megalobrama amblycephala)和翘嘴鲌(Culter alburnus)是中国重要的养殖经济鱼类，分别属于鲤科(Cyprinidae)、鲌亚科(Culterinae)中的鲂属(Megalobrama)和鲌属(Culter)。团头鲂杂食偏草食性，生长速度快，养殖成本低；翘嘴鲌体型细长，肉食性，肉质细嫩鲜美，鳞片薄且细小，易脱落易受伤，生长慢，饲养成本高(农业农村部渔业渔政管理局等, 2020)。虽然二者在食性、生长和抗逆性等性状大不相同，但有很大的性状互补性。杂交可以将双亲优良性状集于一身(周华兴等, 2021)，但这种优势通常只限于F₁代。理论上讲，在三倍体生长和发育期间，由于性腺几乎不发育，可以保持F₁代杂种优势，避免继续繁殖造成的种质衰退，节省的能量将用于加速生长和提高肌肉品质(王雪薇, 2017)，这无疑将大大拓展“鲂鲌”杂交新品种的养殖空间，产生较大的经济、社会和生态价值。

近年来，本课题组开展团头鲂(♀)×翘嘴鲌(♂)杂交三倍体的静水压诱导实验，获得了鲂鲌三倍体[高三倍体率(> 80%)和高成活率(> 98%)]和鲂鲌二倍体，初步观察三倍体具有较强的生长优势。鲂鲌三倍体与二倍体形态类似，外观难以区分，因此，形态分析对种群鉴定与后续选育具有重要意义。目前，开展的杂交实验中与团头鲂、翘嘴鲌相关的形态研究已有记载，如关文志等(2017)对团头鲂、三角鲂(M. terminalis)、长春鳊(Parabramis pekinensis)及其杂交后代共7个群体的形态进行了比较分析；郭洪洪等(2018)对三角鲂、团头鲂、翘嘴鲌及其杂交后代5个群体的形态差异进行了分析；刘加林(2020)对翘嘴鲌和三角鲂亲本及其杂交后代太湖鲂鲌(C. alburnus♀× M. terminalis♂)的形态特征进行了分析；但对经静水压诱导的团头鲂(♀)×翘嘴鲌(♂)杂交后代的形态研究尚未见报道。

本研究通过生长对比实验比较分析鲂鲌三倍体、二倍体及其双亲自交后代4种鱼的生长性能，并运用传统形态学测量法结合现代框架结构测量法对4种鱼的形态进行统计，同时应用多元统计分析法对数据进行分析以探究4种鱼之间的形态差异，旨在为鲂鲌杂交后代种群鉴定及选育提供基础资料。

1 材料与方法 1.1 亲本催产、人工受精及孵化

2021年5月，静水压诱导的团头鲂(♀)×翘嘴鲌(♂)杂交实验在农业农村部团头鲂遗传与育种中心进行。母本为团头鲂“浦江2号”F₆代，父本为淀山湖翘嘴鲌F₃代，均来自农业农村部团头鲂遗传与育种中心，均为性腺发育良好、体格健壮、形体优美的3龄鱼。于傍晚进行催产药物(10 μg HCG+200单位LHRH- A2/kg)注射，雄鱼剂量减半，将亲鱼安置于深1.3 m、半径1 m的圆形产卵池中，保持池中流水状态。翌日清晨，待亲本能挤出卵子和精子时，迅速捞起亲鱼，在避光环境下挤压亲鱼腹部取卵子和精子于盆中，进行人工干法授精，随后用滑石粉进行脱黏处理。经过静水压力(受精后处理起始时间为2~3 min，压强为35~50 MPa，持续时间为3 min)处理后，于孵化桶中孵化受精卵，直至鱼苗可以正常平游再转移到深1.5 m的600 m²大土塘中。从受精、孵化到苗种培育整个过程的水温控制在22~24 ℃。待鱼苗长至3~5 g时通过Partec倍性分析仪区分出鲂鲌三倍体和二倍体，分别用hybrid-3n和hybrid-2n表示。同时建立团头鲂×团头鲂、翘嘴鲌×翘嘴鲌2个自交群体，后代分别用MA和CA表示。

1.2 生长对比

生长对比实验于2022年3月中旬开始，将1.1中4个群体分别记录初始体重W₀并剪鳍条以区分(hybrid-3n：左胸；hybrid-2n：右胸；MA：左腹；CA：右腹)，而后转移至新的深1.5 m的600 m²大土塘中同池混养，整个养殖过程均使用相同的鱼用配合饲料(粗蛋白31.0%、粗纤维12.0%、粗脂肪8.0%)，且每周换水1次保持养殖水体清爽(pH为6.8~7.6，溶解氧 > 5.5 mg/L，氨氮 < 0.5 mg/L)。2022年10月中旬对全池拉网起捕，进行体质量测定以比较生长速度。

绝对增重率AGR(g/d)=(W₁−W₀)/(t₁−t₀)，其中，W₁和W₀分别对应时间t₁和t₀时的体质量，t₁−t₀为210 d。变异系数CV＝标准差/平均值×100%。

1.3 测量方法

2022年12月上旬拉网，在每个群体中都随机取30尾进行计数与测量。对背鳍条、胸鳍条、腹鳍条、臀鳍条、侧线上鳞、侧线下鳞和侧线鳞7项可数性状计数，对体质量、全长(L_a)、体长(L_b)、体高(L_c)、头长(L_d)、吻长(L_e)、眼间距(L_f)、眼径(L_g)、尾柄高(L_h)、尾柄长(L_i)、背棘长(L_j) 11项可量性状与20项框架结构数据(陈杰等, 2014)进行测量(图 1)。数据测定使用电子游标卡尺，精度为0.01 cm。体质量用电子天平，精度为0.1 g。

图 1 框架结构定点图 Fig.1 Fixed point diagram of frame structure 1. 胸鳍起点；2. 吻前端；3. 腹鳍起点；4. 头背部末端；5. 臀鳍起点；6. 背鳍起点；7. 臀鳍未端；8. 背鳍未端；9. 尾鳍腹部起点；10. 尾鳍背部起点。 1. Origin of pectoral fin; 2. Tip of snout; 3. Origin of pelvic fin; 4. Terminus of head back; 5. Origin of anal fin; 6. Origin of dorsal fin; 7. Terminus of anal fin; 8. Terminus of dorsal fin; 9. Ventral origin of caudal fin; 10. Dorsal origin of caudal fin.

1.4 统计分析

为避免鱼体规格对形态参数造成统计误差，将所有可量性状数据转变成比例性状数据，得到9项(郭洪洪等, 2018)，框架结构数据转变成和体长的比例数据。基于可数性状和9项比例性状数据计算团头鲂(♀)×翘嘴鲌(♂)的杂种指数(hybrid index, HI) (Witkowski et al, 1980; Cricelli et al, 1988)。HI=100×(H_i−M_i1)/(M_i2−M_i1)，其中，H_i为杂种平均值；M_i1为母本平均值；M_i2为父本平均值。HI在45~55之间属中间性状，HI > 55为偏父本性状，HI < 45为偏母本性状，HI > 100或者HI < 0为超亲偏离性状。采用SPSS 26对每个可量性状和9个比例性状作单因素方差分析(one-way ANOVA)，LSD法多重比较进行显著性检验，P < 0.05为差异显著。对9个可量比例参数和20项框架比例参数共计29项做聚类分析(欧式距离的最短系统聚类法)、判别分析(逐步判别法)和主成分分析(张尧庭等, 1982)。

2 结果 2.1 形态特征

4种鱼的主要形态特征比较见图 2。仔细观察可以看出，hybrid-3n的形态特征与hybrid-2n较为相似，尤其头部非常相像，皆明显大于双亲自交后代，嘴部皆上翘但不如CA明显。hybrid-3n体长与CA和hybrid-2n都较为相似，但体高明显增加趋向母本MA。hybrid-3n背部略微隆起，较hybrid-2n更为明显，与MA相似，而CA背部较为平整。hybrid-3n背部和侧上方为青灰色，腹部与侧下方为银白色，鳞片较大类似MA，薄软情况类似CA。

图 2 杂种三倍体、杂种二倍体及其双亲自交后代的形态 Fig.2 Morphology of hybrid triploid, hybrid diploid and double inbred offspring

2.2 生长对比

经过210 d饲养，剪去的鳍条清晰可见不存在错分可能，以绝对增重率反映生长速度，hybrid-3n生长速度[(0.88±0.11) g/d]较hybrid-2n提高8.64%，较MA提高20.55%，较CA提高120.00%，表明hybrid-3n更具生长优势(表 1)。

表 1 杂种三倍体、杂种二倍体、团头鲂和翘嘴鲌的生长速度比较 Tab.1 Comparison of growth rates of hybrid triploid, hybrid diploid, M. amblycephala and C. alburnus

2.3 可数性状

4种鱼可数性状表现如表 2所示，可见hybrid-3n和hybrid-2n的臀鳍条、侧线上鳞、侧线下鳞和侧线鳞数4项均介于双亲之间，在侧线上鳞、侧线下鳞和侧线鳞数上，4个群体皆差异显著(P < 0.05)。杂种指数显示，hybrid-3n背鳍和胸鳍为偏父本性状；其他5个性状为偏母本性状。hybrid-3n和hybrid-2n杂种指数平均值分别为41.05和36.07，接近中间值，均略偏向于母本MA。

表 2 杂种三倍体、杂种二倍体及其双亲自交后代的可数性状数据 Tab.2 Countable character data of hybrid triploid, hybrid diploid and their progenies of double inbreeding

性状Character	hybrid-3n	hybrid-3n	MA	CA	HI₁	HI₂
背鳍Dorsal fin	3，8.00±0.26	3，8.03±0.32	3，8.10±0.40	3，8.00±0.00	100.00	70.00
胸鳍Pectoral fin	1，15.23±0.63^b	1，15.77±0.97^a	1，15.73±0.45^a	1，14.83±0.38^b	55.56	–4.44
腹鳍Ventral fin	1，7.90±0.31^bc	1，8.03±0.18^ab	1，7.80±0.41^c	1，8.20±0.41^a	25.00	57.50
臀鳍Anal fin	3，23.63±1.10^b	3，23.10±1.37^b	3，25.00±0.53^a	3，21.03±0.56^c	34.51	47.86
侧线上鳞 Lateral supralinear scale	12.13±0.78^c	12.63±0.61^b	11.63±0.61^d	17.50±0.63^a	8.52	17.04
侧线下鳞 Lateral sublinear scale	10.50±0.51^b	10.04±0.91^c	9.20±0.41^d	12.43±0.77^a	40.25	24.77
侧线鳞Lateral scale	59.53±1.78^c	63.63±1.63^b	53.62±1.35^d	78.77±2.24^a	23.50	39.80
平均值Mean					41.05	36.07
注：HI₁为三倍体杂种指数，HI₂为二倍体杂种指数；背鳍、胸鳍、腹鳍、臀鳍中逗号之前为硬棘数，逗号之后为除硬棘外的鳍条数；表中数据格式为平均值±标准差(n=30)；同一行不同字母表示差异显著(P < 0.05)，下同。 Note: HI₁ is triploid hybrid index, and HI₂ is diploid hybrid index. The number of hard spines in dorsal fin, pectoral fin, ventral fin and anal fin is before comma, and the number of fins except hard spine is after comma. The data format in the table is Mean±SD (n = 30). Different letters in the same row indicate significant differences (P < 0.05), and the same below.

表 2 杂种三倍体、杂种二倍体及其双亲自交后代的可数性状数据 Tab.2 Countable character data of hybrid triploid, hybrid diploid and their progenies of double inbreeding

2.4 可量性状

结果如表 3所示，hybrid-3n群体除全长/体长(L_a/L_b)、体长/尾柄长(L_b/L_i)、尾柄长/尾柄高(L_i/L_h)外，其余6个性状与母本差异显著(P < 0.05)；与父本除体长/头长(L_b/L_d)、头长/眼径(L_d/L_g)外，其余性状皆有显著性差异(P < 0.05)；与hybrid-2n群体只在体长/尾柄长(L_b/L_i)、尾柄长/尾柄高(L_i/L_h)差异不显著(P > 0.05)。其中，在体长/体高(L_b/L_c)、头长/眼间距(L_d/L_f)、体长/背棘长(L_b/L_j)这3个性状上4个群体均有显著性差异(P < 0.05)。

表 3 杂种三倍体、杂种二倍体及其双亲自交后代的可量性状数据 Tab.3 Quantitative character data of hybrid triploid, hybrid diploid and their progenies of double inbreeding

项目Items	hybrid-3n	hybrid-2n	MA	CA	HI₁	HI₂
L_a/L_b	1.08±0.04^a	1.06±0.01^b	1.08±0.01^a	1.06±0.01^b	0	100.00
L_b/L_c	2.80±0.31^c	2.94±0.20^b	2.32±0.04^d	4.56±0.08^a	21.43	27.68
L_b/L_d	4.36±0.23^b	4.50±0.24^a	4.19±0.09^c	4.46±0.05^ab	62.96	114.81
L_d/L_e	3.22±0.17^b	3.36±0.21^a	3.44±0.08^a	2.89±0.06^c	40.00	14.55
L_d/L_g	3.75±0.20^b	3.89±0.21^a	3.32±0.20^c	3.79±0.10^ab	91.84	120.41
L_d/L_f	2.20±0.14^c	2.29±013^b	1.97±0.06^d	2.57±0.07^a	38.33	53.33
L_b/L_i	7.17±0.71^a	7.36±0.89^a	7.36±0.34^a	6.27±0.39^b	17.43	0
L_i/L_h	1.35±0.18^b	1.37±0.11^b	1.28±0.15^b	1.99±0.17^a	9.86	12.68
L_b/L_j	4.92±0.39^c	5.32±0.64^b	4.17±0.31^d	5.71±0.13^a	48.70	74.68
平均值Mean					36.73	57.57
注：L_a：全长；L_b：体长；L_c：体高；L_d：头长；L_e：吻长；L_f：眼间距；L_g：眼径；L_h：尾柄高；L_i：尾柄长；L_j：背棘长。HI₁为三倍体杂种指数，HI₂为二倍体杂种指数。表中数据格式为平均值±标准差(n=30)。 Note: L_a: Full length; L_b: Body length; L_c: Body height; L_d: Head length; L_e: Snout length; L_f: Eye spacing; L_g: Eye diameter; L_h: Caudal stalk height; L_i: Caudal stalk length; L_j: Dorsal spinous length. HI₁ is triploid hybrid index, and HI₂ is diploid hybrid index. The data format in the table is Mean±SD (n=30).

表 3 杂种三倍体、杂种二倍体及其双亲自交后代的可量性状数据 Tab.3 Quantitative character data of hybrid triploid, hybrid diploid and their progenies of double inbreeding

杂种指数显示，hybrid-3n没有表现出超亲偏离，体长/背棘长(L_b/L_j)为中间性状，体长/头长(L_b/L_d)、头长/眼径(L_d/L_g) 2个性状偏向父本，其余6项偏向母本。hybrid-3n和hybrid-2n杂种指数平均值分别为36.73和57.57，显示在可量性状比值上hybrid-3n群体偏向于母本MA，hybrid-2n群体略偏向于父本CA。

2.5 聚类分析

hybrid-3n首先和hybrid-2n聚在一起归为一类，然后再与母本MA聚为一支，最后再与父本CA聚在一起。表明hybrid-3n和hybrid-2n在可量性状和框架结构上受母本的影响要大于父本(图 3)。

图 3 杂种三倍体、杂种二倍体、团头鲂和翘嘴鲌的形态聚类树形图 Fig.3 Morphological clustering tree diagram of hybrid triploid, hybrid diploid, M. amblycephala and C. alburnus

2.6 判别分析

运用逐步判别分析法剔除无显著性差异(P > 0.05)的性状，筛选出11个极显著性差异的生物学性状(P < 0.01)：体长/体高(L_b/L_c)、头长/眼径(L_d/L_g)、L_6~5/L_b、体长/头长(L_b/L_d)、L_4~3/L_b、L_5~7/L_b、头长/吻长(L_d/L_e)、L_4~1/L_b、L_4~5/L_b、L_7~9/L_b、头长/眼间距(L_d/L_f)，分别以X₁~X₁₁表示，构建的判别方程式如下：

hybrid-3n：Y₁＝–3373.274+1687.653X₁–271.121X₂+ 1114.458X₃+5470.034X₄+2226.751X₅+699.818X₆+247.389 X₇+277.876X₈+103.301X₉+65.103X₁₀+260.495X₁₁

hybrid-2n：Y₂＝–3531.189+1626.448X₁–319.090X₂+ 915.873X₃+5872.903X₄+2236.466X₅+610.081X₆+247.127 X₇+288.498X₈+112.040X₉+67.128X₁₀+270.256X₁₁

MA：Y₃＝–3343.923+1843.445X₁–527.639X₂+ 740.523X₃+5804.654X₄+1894.388X₅+1009.327X₆+257.035X₇+268.989X₈+128.266X₉+40.828X₁₀+247.084X₁₁

CA：Y₄＝–3646.496+871.390X₁–233.509X₂+ 1775.813X₃+5563.526X₄+2200.800X₅+1232.343X₆+346.323X₇+247.134X₈+74.288X₉+79.490X₁₀+245.705X₁₁

运用以上判别方程式就可以判断某尾鱼的群体归属，将待测鱼的上述11个形态数据分别代进各判别方程式，结果最大的即为该鱼的归属。判别分析结果见表 4，仅在hybrid-3n和hybrid-2n群体中出现误判，对hybrid-3n和hybrid-2n群体的判别准确率都为90%，hybrid-3n中有3个样本误判为hybrid-2n，hybrid-2n中有3个样本误判为hybrid-3n；综合判别率达95%。

表 4 杂种三倍体、杂种二倍体、团头鲂和翘嘴鲌的判别结果 Tab.4 Discriminant results of hybrid triploid, hybrid diploid, M. amblycephala and C. alburnus

4个群体120个样本的分布如图 4所示，MA和CA群体分别处于图中两端，hybrid-3n和hybrid-2n的群体中心位于父母本之间但更靠近母本MA，且hybrid-3n群体中心相比hybrid-2n要更接近母本。hybrid-3n和hybrid-2n二者所占区域有部分重叠，表明二者在比例性状和框架结构上形态十分接近，不易区分。

图 4 三倍体、二倍体、团头鲂和翘嘴鲌的判别分析图 Fig.4 Discriminant analysis diagram of triploid, diploid, M. amblycephala and C. alburnus

2.7 主成分分析

共获得4个主成分，对变异的累计贡献率为75.10%，占据了群体总变异的大部分。主成分的成分载荷矩阵分析发现，主成分1 (52.87%)主要取决于L6-5/L_b、L8-5/L_b、L5-7/L_b、L6-3/L_b、体长/体高(L_b/L_c)、L4-1/L_b、L6-7/L_b、L2-4/L_b、L4-3/L_b、头长/眼间距(L_d/L_f)、L10-9/L_b、L6-8/L_b、尾柄长/尾柄高(L_i/L_h)、体长/背棘长(L_b/L_j)、头长/吻长(L_d/L_e)、L7-9/L_b和L8-7/L_b，这17项参数指标其载荷绝对值皆大于0.7，主要是对鱼体体高的形态差异表现。对主成分2 (10.06%)起主要作用的参数指标有L1-3/L_b、L8-9/L_b、L3-5/L_b和L4-5/L_b，这4项比例参数载荷绝对值皆大于0.5，主要是对鱼体躯干的形态差异表现。主成分3 (7.08%)和主成分4 (5.09%)分别主要体现了鱼体尾部和头部的形态差异。由此可见，hybrid-3n、hybrid-2n、MA和CA四个群体在形态上的差异主要表现在鱼体体高和躯干处。

3 讨论 3.1 鲂鲌三倍体的生长优势

三倍体鱼类因染色体组数为奇数致使减数分裂出现异常，表现出不育或败育(李宝玉, 2021)，理论上，这将使得原本用于性腺生长发育的能量转而被用来加速个体的生长。Utter等(1983)研究发现，银大马哈鱼(Oncorhynchus kisutch)幼鱼三倍体的生长不如二倍体；Byamungu等(2001)研究发现，奥里亚罗非鱼(Oreochromis aureus)三倍体幼鱼生长速度不如二倍体；Artamonova等(2018)研究发现，细鳞大马哈鱼(O. gorbuscha)三倍体在成鱼时期生长性能明显不如二倍体。Purdom (1976)研究了异源杂交鲽(Pleuronectidae)三倍体和二倍体生长情况，发现二者不具备显著性生长差异；同样，Wolters等(1982)在斑点叉尾 (Ietalurus punetaus)、Hussain等(1995)在尼罗罗非鱼(O. niloticus)、Gillet等(2001)在北极红点鲑(Salvelinus alpinus)研究中也得出相似结论。Soltan等(2017)通过实验发现，尼罗罗非鱼三倍体较二倍体生长快；邹远超等(2017)研究表明，乌鳢(Channa argus)三倍体生长比二倍体快；魏凯等(2019)研究认为，细鳞鲑(Brachymystax lenok)三倍体生长快于二倍体。可见，三倍体鱼类的生长并非完全如理论所说要快于二倍体，其实际情况因鱼而异。我们推测，染色体组数的改变会通过影响性腺发育进而表现出具有高于或低于二倍体的生长速度，其中机制有待进一步研究。Piferrer等(2009)研究发现，多种三倍体鱼类在幼鱼早期的生长速度通常与二倍体相近，在性成熟后生长速度开始高于二倍体。本研究中，hybrid-3n初始体重为(13.84±2.82) g，与其他3种鱼相差不大，经过210 d的养殖实验后，hybrid-3n生长速度较hybrid-2n提高8.6%，较母本MA提高20.5%，较父本CA提高1.2倍，表明鲂鲌三倍体更具生长优势，这与Piferrer等(2009)的研究结果基本一致。因此，鲂鲌三倍体具备在生产上推广应用的基础条件。

3.2 鲂鲌三倍体的形态学变异

形态是生物最直观的外在表现，其差异能直接反映生物多样性，是种群鉴定、分类的重要依据(尹洁等, 2020)。本研究可数性状分析发现，hybrid-3n和hybrid-2n平均杂种指数HI分别为41.05和36.07，均略偏向于母本MA。9项可量比例性状分析发现，hybrid-3n和hybrid-2n的多数可量性状变异较大，hybrid-3n与父本和母本分别有7项和6项存在显著性差异，hybrid-2n有6个比例性状分别与父母本有显著性差异。hybrid-3n和hybrid-2n杂种指数平均值分别为36.73和57.57，显示hybrid-3n可量性状比值偏向于母本，hybrid-2n可量性状比值偏向于父本，这与上述可数性状结果略有不同，可能是因为可量性状为连续性状，受环境影响更大，不如可数性状稳定(宋文, 2016)。

表 5 4个群体的主成分载荷矩阵及主成分的贡献率 Tab.5 Principal component load matrix and principal component contribution rate of four groups

项目Items	主成分Principal component
项目Items	1	2	3	4
L_6~5/L_b	0.978	0.058	–0.048	–0.001
L_8~5/L_b	0.975	0.055	–0.035	–0.010
L_5~7/L_b	0.972	–0.014	–0.002	–0.056
L_6~3/L_b	0.970	0.033	–0.072	–0.011
L_b/L_c	–0.965	–0.047	0.116	–0.057
L_4~1/L_b	0.929	0.123	0.160	–0.009
L_6~7/L_b	0.907	0.191	–0.146	0.015
L_2~4/L_b	0.900	–0.090	0.254	–0.050
L_4~3/L_b	0.897	0.275	–0.052	0.081
L_d/L_f	–0.887	0.114	0.094	0.153
L_10~9/L_b	0.866	0.125	0.200	–0.011
L_6~8/L_b	0.862	0.168	0.225	–0.156
L_i/L_h	–0.848	–0.085	0.320	–0.025
L_b/L_j	–0.817	0.139	–0.285	–0.167
L_d/L_e	0.800	0.274	–0.032	0.060
L_7~9/L_b	–0.794	0.188	0.288	–0.016
L_8-7/L_b	0.749	0.305	–0.264	0.006
L_1-2/L_b	0.681	0.016	0.353	0.077
L_b/L_d	–0.504	0.343	–0.455	–0.491
L_d/L_g	–0.487	0.437	–0.263	0.408
L_1~3/L_b	–0.018	0.713	0.033	–0.024
L_8~9/L_b	0.195	0.636	0.316	–0.219
L_3~5/L_b	–0.460	0.589	–0.040	0.176
L_4~5/L_b	–0.202	0.509	–0.331	0.466
L_4~6/L_b	–0.220	0.500	0.083	0.291
L_8~10/L_b	–0.281	0.473	0.229	–0.269
L_10~7/L_b	–0.239	0.364	0.627	–0.221
L_b/L_i	0.526	0.010	–0.599	–0.217
L_a/L_b	0.325	–0.226	0.234	0.646
贡献率Contribution/%	52.87	10.06	7.08	5.09

表 5 4个群体的主成分载荷矩阵及主成分的贡献率 Tab.5 Principal component load matrix and principal component contribution rate of four groups

现今，已有众多的物种鉴定、育种及进化方面的研究都综合利用了各多元统计分析方法进行形态差异比较(梁述章等, 2018; 朱书琴等, 2021; 周华兴等, 2021; 马凯等, 2023)。本研究聚类分析显示，hybrid-3n首先和hybrid-2n聚在一起归为一类，然后与母本MA聚为一支，最后与父本CA聚在一起，表明hybrid-3n和hybrid-2n在可量性状和框架结构上更接近母本，表现出一定母性效应，类似于杂交种“云龙石斑鱼” (Epinehelus moara ♀ × E. lanceolatus ♂) (李振通等, 2019)、大口黑鲈(Micropterus salmoides♀)×蓝鳃太阳鱼(Lepomis mearchirus♂)杂交F₁ (李武辉等, 2020)和团头鲂(♀)×长春鳊(♂)杂交F₁ (赵博文等, 2015)等大多数杂交后代的整体形态表现。但也有杂交后代形态偏向父本的报道，如草鱼(Ctenopharyngodon idella♂) ×赤眼鳟(Spualiobarbus curriculus♀) F₁ (金燮理等, 1999)和翘嘴红鲌(Erythroculter ilishaeformi♀)×团头鲂(♂) F₁的形态特征(顾志敏等, 2008)。这种差异可能是母性效应在不同性状中表现不同，具体原因有待进一步探索。

本研究判别分析对hybrid-3n和hybrid-2n群体的判别准确率均为90%，对MA和CA判别准确率均为100%，4种鱼的综合判别率达95%，判别效果较好。可见hybrid-3n和hybrid-2n因形态相似，不可避免出现误判。判别分析散点图显示，MA和CA两群体分别占据不同区域，hybrid-3n和hybrid-2n的分布中心位于父母本之间且更靠近母本MA；hybrid-3n和hybrid-2n二者所占区域有部分重叠，表明二者在比例性状和框架结构上形态十分接近，不易区分。这些与聚类分析所得结果基本一致。构建判别方程式的11个生物学性状大多与身体纵轴相关尤其体现在与体高的比例上，躯干处也有涉及，这与主成分分析所得结果类似。

本研究主成分分析共获得4个主成分，累计贡献率为75.10%，可以较好概括4种鱼群体间的形态差异。其中，决定主成分1 (52.87%)特征向量绝对值较大的性状大多体现在与体高的比例上，这与易伯鲁(1955)的研究结果相符。此外，主成分2 (10.06%)中决定它的4项参数主要集中在鱼体躯干处，反映鱼体躯干处的形态特征也是鱼类变异的重要来源。何震晗(2021)对不同地理种群的黄鳍棘鲷(Acanthopagrus latus)进行主成分分析认为，它们的差异主要体现在鱼体躯干和尾部性状上；张秀霞等(2017)研究发现，唐鱼(Tanichthys albonubes)形态变异主要在眼径大小和头部；王新月等(2022)认为不同地理群体的叶尔羌高原鳅(Triplophysa yarkandensis)形态差异主要在机体头部和躯干前段。这些与本研究所得结果有异，可能是因为鱼类尾部是行进的重要器官，与游泳能力密切相关，鱼类头部与食性摄食密切相关；而躯干和体高可以反映是否有足够空间积累脂肪增加体重；另外，蔡鸣俊等(2001)认为，较大的体高可以减轻被凶猛鱼类吞噬的危险。本研究的实验鱼为静水池塘养殖，饵料丰富，与上述鱼类生活环境大不相同，因此，研究结果的异同可能是由于鱼类对生态环境的适应。

可见，本研究所用3种多元分析方法有效地从不同角度反映了hybrid-3n、hybrid-2n、MA和CA四个群体间的形态差异，这使得彼此之间不可替代。生物形态由遗传因子与环境共同决定(杨慧, 2016)，本研究中4种鱼均同池混养，养殖环境相同，造成的形态差异可能更多的是遗传因子的反映。我们通过对4种鱼进行染色体核型分析认为hybrid-2n从母本和父本中各继承1n染色体，因母性效应而使形态偏向于母本，而hybrid-3n因静水压力对卵子第二极体排出的抑制从母本继承了2n染色体，从父本继承1n染色体而使得形态较hybrid-2n与母本更为接近。

综上所述，本研究证实了鲂鲌三倍体具有优越的育种潜力，并为鲂鲌杂交子代的形态比较提供基础数据对鲂鲌鱼类种群资源的鉴定保护具有积极意义，有望为优良鲂鲌新品系的建立提供理论基础。

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