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Acta Agronomica Sinica ›› 2021, Vol. 47 ›› Issue (6): 1043-1053.doi: 10.3724/SP.J.1006.2021.04069


Creation of male sterile germplasm using the partial length gene of HcPDIL5-2a in kenaf

ZHOU Bu-Jin1,2(), LI Gang3, JIN Gang4, ZHOU Rui-Yang1,*(), LIU Dong-Mei5, TANG Dan-Feng3, LIAO Xiao-Fang4, LIU Yi-Ding1, ZHAO Yan-Hong4, WANG Yi-Ning1   

  1. 1College of Agriculture, Guangxi University, Nanning 530004, Guangxi, China
    2Maize Research Institute of Guangxi Academy of Agricultural Sciences, Nanning 530007, Guangxi, China
    3Guangxi Botanical Garden of Medicinal Plants, Nanning 530023, Guangxi, China
    4Cash Crop Institute of Guangxi Academy of Agricultural Sciences, Nanning 530007, Guangxi, China
    5Shangqiu Normal University, Shangqiu 476000, Henan, China
  • Received:2020-03-17 Accepted:2021-01-13 Online:2021-06-12 Published:2021-01-22
  • Contact: ZHOU Rui-Yang E-mail:zhou_bujin@163.com;ruiyangzhou@aliyun.com
  • Supported by:
    The National Natural Science Foundation of China(31571719);The Natural Science Foundation of Guangxi(2018JJB130072)


In order to create transgenic male sterile germplasm in kenaf, the partial length of gene HcPDIL-2a was transformed into maintainer line 722HB via pollen tube pathway. A low-temperature sensitive male sterile mutant called ‘722THS’ was accquired, which showed sterile when grown in winter in Hainan and fertile when grown in summer in Nanning, and a stable Genic Male Sterile (GMS) line 722HS was selected from its sister progenies. A cytoplasmic male sterile line 722HA was bred by crossing the mutant 722THS with the wild type 722B. Cytological observation indicated that the microspore abortion of 722THS and 722HA both happened at dinuclear stage. The mitochondrial DNA molecular tag revealed that mitochondrial DNA rearrangement occurred in 722HA, while mitochondrial DNA of 722THS and 722B remained unchanged. This study provided a new way for the creation of male sterile germplasms, which had important scientific significances and broad application prospects.

Key words: kenaf, male sterility, partial length HcPDIL-2a

Fig. 1

Plasmid vector map of PBI121-PH-GFP"

Fig. 2

Morphological characteristics of the flowers from 722THS, 722HB, and 722A in kenaf"

Fig. 3

Microspore development observation of mutant 722THS and wild type 722B in kenaf"

Table 1

Fertility statistics of 722THS/722B hybrid F1 and F2"

Number of fertile plants
Number of sterile plants
722THS-1/722B 86 0
722THS-2/722B 113 0
(722THS-1/722B)自交 (722THS-1/722B) selfing 130 0
(722THS-2/722B)自交 (722THS-2/722B) selfing 95 0

Fig. 4

PCR detection of 722THS natural hybrid F1 sterile plants M: DNA marker; 1: positive plasmid; 2: negative control; 3-6: sterile plants."

Fig. 5

NPT II detection of fertile self-progeny after girdling of 722THS in Nanning of Guangxi M: DNA marker; 1-6: sterile plants; 7-31: fertile plants."

Table 2

Fertility statistics of fertile self-progeny after girdling of 722THS in Hainan"

Count date
temperature (℃)
Hybrid combination
Number of fertile plants
Number of sterile plants
Expect proportion
χ2 显著性测验
Significance test
χ2 (0.05,1)
Early flowering (from March 14
to 16)
23.3 722THS-S1环割可育自交
722THS-S1 girdling fertile selfing
123 91 3:1 35.05 NS
722THS-S2环割可育自交722THS-S2 girdling fertile selfing 263 193 3:1 72.99 NS
722THS-S3环割可育自交722THS-S3 girdling fertile selfing 133 101 3:1 41.17 NS
722THS-S4环割可育自交722THS-S4 girdling fertile selfing 17 44 3:1 61.55 NS
Late flowering
(from April 11
to 13)
27.5 722THS-S1环割可育自交
722THS-S1 girdling fertile selfing
158 56 3:1 0.24 *
722THS-S2环割可育自交722THS-S2 girdling fertile selfing 324 122 3:1 1.50 *
722THS-S3环割可育自交722THS-S3 girdling fertile selfing 163 71 3:1 3.56 *
722THS-S4环割可育自交722THS-S4 girdling fertile selfing 43 18 3:1 0.66 *

Fig. 6

Morphological characteristics of the flowers from 722HA and 722HB in kenaf [23]"

Fig. 7

Microspore development observation of 722HA and 722HB in kenaf [23] PMC: pollen mother cell; Td: tetrad microspore; Ms: mononuclear microspore; Ds: dinuclear microspore; MP: mature pollen grain; Ep: epidermis; En: endothecium; ML: middle layer; T: tapetum; Bar: 20 μm."

Fig. 8

Cytoplasmic types detection of different materials using molecular tags MM509 and MM556 M: DNA marker; 1, 2, 3, 4 represent 722A, 722HA, 722B, and 722THS, respectively."

Fig. 9

Sequence alignment of molecular tags MM509 and MM556 in different materials A: sequence alignment with molecular tag MM509; B: sequence alignment with molecular tag MM556."

Table 3

Fertility statistics of hybrid progenies from kenaf CMS line 722HA and 722A with restoring lines"

Number of fertile plants
Number of sterile plants
722A/福红992 722A/Fuhong 992 56 0
722A/R7 91 0
722HA/福红992 722HA/Fuhong 992 52 14
722HA/R7 44 35
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