红心柚和美国枸橼杂种后代对柑橘溃疡病的抗性评价

doi:10.16420/j.issn.0513-353x.2022-0094

摘要/Abstract

摘要：

为了探究枸橼对柑橘溃疡病（Xanthomonas citri subsp. citri，Xcc）的抗性遗传规律，以感溃疡病品种红心柚为母本，与抗溃疡病种质美国枸橼进行杂交，获得了F₁代杂种，并以F₁自交后获得了F₂代。对F₁杂种进行活体叶片注射接种鉴定，对所有F₂代253株进行离体叶片注射接种鉴定，随后对其中的225株F₂活体叶片进行注射接种鉴定，并对部分F₂代活体叶片接种后进行溃疡病菌定量分析。鉴定结果表明，红心柚和美国枸橼杂交获得的75个F₁全部为感病，没有抗感性状分离。对253株F₂植株离体叶片鉴定结果显示，76株表现为抗病，177株表现为感病。对225株F₂植株活体叶片鉴定结果显示，72株为感病，症状与感病品种‘冰糖橙’类似；118株为较感病，发病程度较‘冰糖橙’轻，与感病亲本红心柚的病症相似；10株为较抗病，抗性接近于抗病亲本美国枸橼；4株为抗病，抗性与抗病种质枸橼C-05相似；此外有21株接种溃疡病后，叶片正、反面感病症状差异较大，10株叶正面感病，反面抗病，11株叶反面抗病，叶正面感病。溃疡病菌菌定量与活体鉴定的结果相吻合。综上所述，F₁杂种没有出现抗感性状分离，F₂代植株存在抗感性状分离，抗感比例为1︰15，表明抗感性可能是数量性状遗传。

关键词: 枸橼, 红心柚, F₂代植株, 柑橘溃疡病, 抗性鉴定, 溃疡病菌定量

Abstract:

In order to investigate the genetic mechanism of citron in resistance against citrus canker disease（Xanthomonas citri subsp. citri，Xcc），in this study，the susceptible variety Red pomelo was crossed with the resistant genotype American citron and F₁ hybrids were obtained. F₂ progeny was further gained by self-crossing the F₁ plants. Resistance identification wan performed on the F₁ hybrid plants by in vivo inoculation of Xcc. All the F₂ plants were in vitro inoculated and 225 F₂ plants were successively identified for the resistance by in vivo inoculation. Bacterial quantification was conducted on some of the F₂plants following the inoculation with Xcc. The results from in vivo inoculation of F₁ plants indicated that all F₁ hybrids were susceptible，and no segregation between resistance and susceptibility was observed on the F₁ progeny. In vitro identification of 253 F₂ plants showed that 76 plants were resistant and 177 were susceptible. The results of in vivo inoculation on 225 F₂ plants demonstrated that 72 plants were susceptible，with symptoms similar to those of the susceptible variety Bingtang sweet orange，118 slightly less susceptible than Bingtang sweet orange，but similar to those of the susceptible parent Red pomelo；10 were resistant，similar to the resistant parent American citron and four were resistant as the resistant genotype citron C-05. In addition，the symptoms of 21 F₂ plants showed different reaction following the inoculation with Xcc between the adaxial and abaxial leaf surface. On 10 plants，typical canker symptoms appeared on the adaxial leaf surface but not on the abaxial one；while for the other 11 plants，the typical symptoms inversely emerged on the abaxial leaf surface and not on the adaxial leaf surface. The quantification results of Xcc on the inoculated leaves was consistent with the of in vivo identification. In summary，no segregation of resistance and susceptibility was observed in F₁ progeny，but did in the F₂ generation. The ratio of resistant to susceptible F₂ plant was 1︰15，indicating that resistance and susceptibility may be inherited as quantitative traits.

Key words: citron, Red pomelo, F₂ plant, citrus canker disease, resistance identification, quantification of Xcc

中图分类号:

S666

王萍, 盛玲, 杨锦鹏, 周凌磊, 金燕, 罗旭钊, 马先锋, 邓子牛. 红心柚和美国枸橼杂种后代对柑橘溃疡病的抗性评价[J]. 园艺学报, 2023, 50(4): 765-777.

WANG Ping, SHENG Ling, YANG Jinpeng, ZHOU Linglei, JIN Yan, LUO Xuzhao, MA Xianfeng, DENG Ziniu. Evaluation of Resistance to Citrus Canker Disease in Hybrid Progeny of Red Pomelo and American Citron[J]. Acta Horticulturae Sinica, 2023, 50(4): 765-777.

图/表 9

图1 柑橘植株活体叶片接种溃疡病菌示意图

Fig. 1 Schematic diagram of in vivo inoculation of Xcc

图2 F2柑橘活体叶片接种溃疡病菌后症状分级及代表单株

Fig. 2 F2 citrus live leaves are graded and represent individual strains after inoculation with canker bacteria

图3 红心柚与美国枸橼杂交F1部分单株活体叶片注射接种（105和104 cfu · mL-1）溃疡病菌后（20 d）的症状

Fig. 3 Symptoms at 20 d after injection of single live leaf of F1 part of Red pomelo and American citron （105 and 104 cfu · mL-1）with ulcerative pathogen

图4 红心柚与美国枸橼杂交F2部分单株离体叶片接种（105、106和107 cfu · mL-1）溃疡病菌后（10 d）的症状

Fig. 4 Symptoms of single isolated leaves of hybrid F2 of Red pomelo and American citron inoculated with ulcerative pathogen（105，106 and 107 cfu · mL-1）at 10 d

表1 红心柚 × 美国枸橼F2代活体接种溃疡病症状等级分类

Table 1 Classification of canker disease sumptoms following in vivo inoculation of F2 plants of Red pumelo × American citron

F₁ 单株 F₁ plant	植株数 Number of plants
F₁ 单株 F₁ plant	F₂	等级1 Grade Ⅰ	等级2 Grade Ⅱ	等级3 Grade Ⅲ	等级4 Grade Ⅳ	叶正面极感 Adaxial leaf surface sensation	叶反面极感 Abaxial leaf surface serious
E48	156	3	5	70	46	5	9
E73	63	1	1	31	23	1	1
E65	16	—	22	7	—	4	—
E5	18	—	2	10	3	—	1
总计（株） Total（plant）	253	4	10	118	72	10	11

图5 红心柚与美国枸橼杂交的F2代部分单株活体叶片接种溃疡病菌（105和104 cfu · mL-1）后（20 d）的症状

Fig. 5 Symptoms（20 d）of single living leaves of the F2 part of a hybrid of Red pumelo × American citron inoculated with ulcerative bacteria（105 and 104 cfu · mL-1）

图6 红心柚与美国枸橼杂交的F2代部分单株活体接种溃疡病菌（105和104 cfu · mL-1）后（20 d）叶正、反面差异大的症状

Fig. 6 Symptoms of large difference between the front and back sides of the leaves（20 d）of a single plant of F2 hybridization of Red pumelo × American citron inoculated with ulcerative bacteria（105 and 104 cfu · mL-1）

图7 不同抗性种质与部分F2植株叶片接种后溃疡病菌的生长量 BTC：‘冰糖橙’；HXY：红心柚；JYC05：枸橼C05；MGJY：美国枸橼。不同小写字母表示不同叶片接种后溃疡病菌含量差异显著（P < 0.05）。

Fig. 7 Growth of ulcerative bacteria after inoculation with different resistant germplasm and some F2 plant leaves BTC：‘Bingtang’sweet orange；HXY：Red pommelo；JYC05：Citron C-05；MGJY：American citron. Different lowercase letters indicate signicant differences in the content of ulcerative bacteria between different leaves（P < 0.05）.

表2 红心柚 × 美国枸橼F2后代植株对柑橘溃疡病抗感情况汇总

Table 2 Summary of the resistant and susceptle F2 plants of Red pomelo × American citron

F₁单株 F₁plant	植株数 Number of plants
F₁单株 F₁plant	F₂	活体鉴定 Identified	抗病株 Resistant plant	感病株Susceptible plant
E48	156	138	8	130
E73	63	58	2	56
E65	16	13	2	11
E5	18	16	2	14
总计Total	253	225	14	211

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