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African Journal of Agricultural Marketing ISSN 2375-1061 Vol. 11 (1), pp. 001-007, January, 2023. Available 
online at www.internationalscholarsjournals.org © International Scholars Journals 

 

Author(s) retain the copyright of this article. 
 
 

Full Length Research Paper 

 

Use of male gametocide: An alternative to 

cumbersome emasculation in coriander 

(Coriandrum sativum L.) 
 

Giridhar Kalidasu1*, C. Sarada1, P. Venkata Reddy1 and T. Yellamanda Reddy2
 

 
1
Horticultural Research Station, Lam, Guntur-522 034, Andhra Pradesh, India. 

2 Planning and Monitoring Cell, ANGRAU, Rajendranagar, Hyderabad-500 030, Andhra Pradesh, India 
 

Accepted 12 October, 2022 
 
New varieties of the coriander are traditionally bred using either mass selection or pure line selection or recurrent 
selection. Attempts to use hybridization to combine desirable traits were scarcely successful due to cumbersome 
emasculation. Hence, using male gametocides as an alternative to hand emasculation offers immense scope. The 
present study is taken up to assess five gametocides viz. Gibberellic acid, 2, 4-D, Maleic Hydrazide, Ethrel and Surf 
Excel at different concentrations which were sprayed at the time of flower primordia initiation. Pollen viability and 
pollen germination were assessed periodically. The results indicated that none of the gametocides evaluated resulted 
in pollen sterility and pollen germination was unaffected. However, spraying of Maleic Hydrazide at 125 ppm resulted in 
suppression of anther dehiscence due to severe agglutination of the pollen. Repeat spraying of Maleic Hydrazide at 
100 ppm from 25 DAS on wards until the cessation of flowering caused suppression of anther dehiscence during entire 
flowering period. Successful crosses were obtained using Maleic Hydrazide as chemical emasculation agent. 
 

Key words: Coriander, coriandrum, emasculation, male gametocide, crossing, hybridization. 

 
INTRODUCTION 

 
Coriander is a facultative cross pollinated crop. The 
inflorescence is a compound umbel. Peripheral florets of the 
umbellets are hermaphrodite and central florets are 
staminiferous or sometimes sterile (Diederichsen, 1996). 
Romanenko et al. (1991) showed that plants that were not 
emasculated but were pollinated with pollen of other plants 
still had a degree of selfing of 25%. Crossing in coriander is 
acknowledged as cumbersome due to difficult process of 
emasculation (Romanenko et al., 1992). Hence, new 
varieties of the crop are traditionally bred using either mass 
selection or pure line selection or recurrent selection or 
through mutation breeding. Attempts to use hybridization to 
combine desirable traits were scarcely successful due to 
inherent disadvantages in the crop for successful 
emasculation and crossing. Hence, using male gametocides 

as an alternative to hand emasculation offers immense  
 
 

 
*Corresponding author. E-mail: gkalidasu@yahoo.com. 
 
Abbreviations: MH- Maleic Hydrazide (1, 2-dihydropyridazine, 

3-6-dione). GA - Gibberellic Acid. 2, 4-D- 2, 4-dichloro phenoxy 

acetic acid. PGM- Pollen Germination Medium. 

 
 
 

 
scope for development of new varieties. 

Several gametocides have been reported effective in 
inducing pollen sterility in various crops. Sodium methyl 
arsenate, 2,3-dichloroisobutyrate, sodium 2,2-dichloro-
propionate, Gibberellic acid, Maleic Hydrazide (1,2-
dihydropyridazine, 3-6-dione), 2,4-dichloro phenoxy 
acetic acid, ethyl 4-fluorooxanilate, Trihalogenated 
methylsulfonamides, ethyl and methyl arsenates, and 
many other chemicals were reported to have male 
gametocide effects in several crops. Salgare (2004) 
reported that foliar application of all the concentrations of 
maleic hydrazide above 50, 200, 800 µg/ml suppressed 
cent per cent pollen germinability of Phaseolus mungo, P. 
aureus, Cyamopsis tetragonoloba, respectively showing 
prospect for use as male gametocide. Garcia Torres et al. 

(1979) have shown that GA3 150 ppm induced of 

maximum pollen sterility in sunflower. Lakshmi Praba and 
Thangaraj (2005) reported that ethrel (800 ppm), salicylic 
acid (600 ppm) and maleic hydrazide (0.2%) induced a 
significantly higher percentage of male sterility in the 
TGMS lines of rice. Singh (1999) in rice, Chauhan and 
Vandana Singh (2002) in mustard and Gangaprasad et 
al. (2004) in Niger (Guizotia abyssinica) reported induction 



2 

 

  
 
 

 
Table 1. The composition of the Pollen Germination medium for each 1000 ml 

of water.  
 

 S/No Ingredient Quantity 

 1. Sucrose 100 g 

 2. Boric acid (H3BO3) 100 mg 

 3. Calcium nitrate (Ca (NO3)2. 4H2O) 300 mg 

 4. Magnesium Sulfate Heptahydrate (MgSO4. 7H2O) 200 mg 
 5. Potassium nitrate (KNO3) 100 mg 

 

 

of very high pollen sterility at concentrations of one to six 
per cent of detergent.  

Due to its herbaceous nature of coriander with small 
and delicate flowers, the crop is sparsely amenable for 
crossing. In view of this, the present study is taken up to 
assess five male gametocides viz. Gibberellic acid, 2, 4 - 
D, Maleic Hydrazide, Ethrel and Surf Excel at different 
concentrations using Sadhana as test variety. 
 

 
MATERIALS AND METHODS 
 
The experiments were conducted during Rabi seasons of 2007 - 

2008 and 2008 - 2009. 

 

Chemicals evaluated and test variety 
 
Five gametocides viz. Gibberellic acid, 2, 4-D, Maleic Hydrazide, 
Ethrel and Surf Excel Quick Wash (active ingredient: sulphonated 
methyl ester) were evaluated. GA at 50, 100, 150 and 300 ppm; 2, 
4-D at 10, 50, 100 and 500 ppm; Maleic Hydrazide at 50, 75, 125 
and 250 ppm; Ethrel at 1000, 2000, 3000 and 5000 ppm; Surf Excel 
at 0.25, 0.5, 0.75 and 5% and distilled water spray as control were 
used. The test variety chosen was Sadhana which is a medium 
duration variety with 85 - 100 days duration. The variety comes to 
50% flowering between 45 - 55 days. The variety shows flower 
primordia initiation between 30 - 40 days. All the gametocides were 
sprayed once at the time of flower primordia initiation using uniform 
quantity of spray fluid of 22 ml per one meter length of plant row. 
Three meter lengths of rows containing 30 plants were used for 
applying the gametocides. While care was taken so that all the 
plants in the row are evenly sprayed. Immediately after spraying, 
twenty plants were randomly selected and labeled for subsequent 
observations. 

 

Pollen collection, assessment of pollen sterility and pollen 

germination 
 
In all the treatments fresh pollen was collected in the field from 
recently opened anthers showing fresh pollen, from the florets of 
the treated plants. While collecting the pollen, care was taken so 
that the pollen was from at least five different plants. The pollen was 
mixed to make a single lot. A tiny droplet of the 1% acetocarmine 
stain was placed on the slide. Using a teasing needle, a small 
amount of pollen of this lot was placed on stain and thoroughly 
mixed to ensure uniform penetration of the stain into the pollen. 
Cover slips were gently placed on to different slides for each 
treatment. The slides were then observed under a microscope.  

The stained pollen was examined under microscope and at least 

five hundred pollen grains were counted in each sample by select-

ing random fields. Tests were repeated whenever there was a full 

 
 
staining of all the pollen grains or partial dying was observed. Tests 
were initiated on the seventh day of the spraying and repeated for 
five times with an interval of two days. Pollen sterility estimated thus 
was converted to percentage from the number of non-dyed and 
total pollen.  

In vitro pollen germination was assessed using Pollen 
Germination Medium (PGM). The PGM was prepared using Brew-
baker and Kwack’s (1963) preparation method. The composition of 
the medium for each 1000 ml of water is presented in Table 1. The 
medium was prepared from dissolving the above ingredients in one 
litre of distilled water.  

Freshly collected pollen was spread on a cover slip and was 
placed on a slide having a tiny droplet of PGM solution. The slides 
were placed in an incubator at 25°C. Each treatment was assessed 
for germination after three and six hours of incubation. A minimum 
of 100 pollen grains were examined for each observation. 
Germination frequencies were recorded by counting germinated 
and non-germinated pollen grains. Pollen grains with development 
of pollen tube (germinated) and without are counted, and 
germination was expressed in percentage.  

Observations on anther dehiscence were also recorded using a 

stereomicroscope. The specific findings on anther dehiscence were 
further verified using another test variety Swathi by spraying Maleic 
Hydrazide at 125 ppm on fifty pre-labelled plants. 

 

Phytotoxicity and related plant growth assessment 
 
Phytotoxicity was assessed periodically starting from the second 

day of the spraying with seven days interval. Toxicity symptoms on 

leaves, stem, and inflorescence were recorded. 

 
Evaluation of MH use in chemical emasculation 
 
The effect of maleic hydrazide in preventing the pollen dispersal 
through pollen agglutination which was observed during first year of 
testing was further evaluated for its use as chemical emasculation 
agent in test variety Sudha. Three spray schedules, that is. maleic 
hydrazide 100 ppm at weekly interval from 25 DAS to cessation of 
flowering, maleic hydrazide 125 ppm at weekly interval from 25 
DAS to cessation of flowering and maleic hydrazide 100 ppm at 25 
DAS followed by maleic hydrazide at 125 ppm subsequently at 
weekly interval to cessation of flowering were tested to compare the 
efficacy of maleic hydrazide in sustaining the pollen agglutination. 
Pollen agglutination from the treated samples was monitored 
periodically through the use of stereomicroscope during post-spray 
period until the cessation of flowering. Bagging of umbels of treated 
plants was carried out to observe any seed set for confirming the 
visual observations. 

 
Assessment of Maleic Hydrazide use in chemical emasculation 
 
The maleic hydrazide use in breeding programmes was assessed 



3 

 

 
 
 

 
Table 2. Phytotoxicity symptoms observed on treated plants.  

 
 Chemical Phytotoxicity Symptoms 

 

 GA 50 ppm No phytotoxicity. 
 

 GA 300, 150 and 100 ppm Elongation of plant and inflorescence. Plants are tender and weak. 
 

 2,4-D 10 ppm No pronounced phytotoxicity. 
 

 2,4-D 50 ppm Similar symptoms as above to a lesser extent. 
 

  Severely convoluted plants. Stem thickens showing some pink pigmentation. Leaves elongated 
 

 
2,4-D 100 ppm 

and drooping. Thickened leaves. Distortion of leaf shape from normal, showing less pinnation 
 

 and uneven pinnation. Abnormal thickening and shape of umbel and umbellets. Abnormal floret  

  
 

  shape. Shortening of flower stalk. Higher number of male flowers in umbels. 
 

 2,4-D 500 ppm Mortality followed by the symptoms described for 2, 4-D 100 ppm below. 
 

 
MH 125, 75 and 50 ppm 

Reduction in plant growth and foliage. Reduction in number of umbellets and hermaphrodite 
 

 florets.  

  
 

 
MH 250 ppm 

Scorching of stem and leaves. Shortening of plant and reduction in plant growth. Reduction in 
 

 foliage size and number. Increased number of umbels but reduced number of umbellets. 
 

  Reduction in number of umbellets and hermaphrodite florets. 
 

 
Ethrel 2000 and 1000 ppm 

Chlorosis of leaves with occasional drying of older leaves. Stem pigmentation and slenderness. 
 

 Early and higher pinnation of leaflets. General growth suppression.  

  
 

 
Ethrel 5000 and 3000 ppm 

Severe chlorosis of leaves. Drying of older leaves. Stem pigmentation and slenderness. Early 
 

 and higher pinnation of leaflets. General growth suppression. Early senescence.  

  
 

 Surf Excel 1.0, 0.75 and 0.5 % No pronounced phytotoxicity. 
 

 Surf Excel 5.0 % Scorching of leaves on the edges. 
 

 

 
using seven male parents and two female parents. The female 
parents were sprayed with maleic hydrazide 100 ppm at weekly 
interval from 25 DAS onwards until the end of flowering cessation. 
The pollen agglutination was monitored through the use of 
stereomicroscope periodically. Further, crossing was taken up by 
dusting of pollen of selected male parents on umbels of female 
parents. Twenty umbels were used for each cross. Number of 
hermaphrodite flowers was recorded before the pollination. Repeat 
pollination was taken up for four consecutive days to ensure inner 
florets also received pollen from selected male parent. Such 
crossed umbels were bagged, tagged and assessed for seed set. 
Selfing was recorded in both maleic hydrazide treated and 
untreated female parents. 

 

 

RESULTS AND DISCUSSION 

 

Male gametocides were found quite effective in various 
crops. However, there is a paucity of information on the 
use of male gametocides in coriander. The present 
investigation threw several insights on the use of male 
gametocides in coriander. 

 

 

Phytotoxicity on treated plants 

 

Among the gametocides evaluated, GA at 50 ppm, 2, 4-D 
at 10 ppm and Surf Excel at 0.25, 0.75 and 1.0% did not 
show any symptoms of phytotoxicity. All other concentra-
tions of the chemicals evaluated caused phytotoxicity 
(Table 2). Most pronounced was the effect of 2, 4-D at 
500 ppm that caused severe phytotoxicity and plant 
mortality within three weeks of spraying. Salgare (1999) 

 

 

who worked on P. mungo for induction of male sterility 

using 2, 4-D, reported that 2,4-D concentration ranging 
from 200 to 5000 g per ml caused mortality of all plants, 
which is similar to the present study. Plants in all the 

treatments except for 2, 4-D 500 ppm, completed life 
cycle with a distinct flowering phase. 
 

 

Pollen sterility and pollen germination 

 

Among the gametocides studied none have shown any 
significant gametocidic effect on pollen except for 2, 4-D. 
Pollen sterility among the treatments ranged from 0 - 
23.2%. The chemicals Gibberellic acid, Maleic hydrazide, 
Ethrel and Surf Excel at lower concentrations showed 
less than 3% sterility. However, 2, 4-D at 100, 50 ppm 
and maleic hydrazide at 250 ppm showed 23.2%, 6.2 and 
8.5% of pollen sterility respectively (Table 3).  
Only 2, 4-D at 100 ppm showed considerable degree of 
pollen sterility (23.2%). However, the sterility recorded 
with 2, 4-D at 100 ppm was not continuous but with a 
peak at fifteen days after spraying. Rustagi and Mohan 
Ram (1971) reported the occurrence of rhythms of pollen 
non-viability interspersed with periods of restoration of 
viability with the use of Dalapon and Mendok in Linseed. 
Salgare (2004) reported that maleic hydrazide at 800 
µg/ml failed to suppress cent per cent pollen fertility in P. 
mungo, P. aureus, Cyamopsis tetragonoloba and Vigna 
mungo.  

Observations on pollen germination showed cent 

percent germination in all the treatments tried except for 



4 

 

 
 

 
Table 3. Pollen sterility (%) and pollen germination (%) in treated plants.  
 
 Chemical % % pollen germination** Chemical % sterility* % pollen 

  sterility *    germination ** 

 GA 300 ppm 0 100.0 MH 50 ppm 2.5 100.0 

 GA 150 ppm 1.2 92.7 Ethrel 5000 ppm 2.9 100.0 

 GA 100 ppm 0.7 100.0 Ethrel 3000 ppm 0.9 100.0 

 GA 50 ppm 0.5 100.0 Ethrel 2000 ppm 0 100.0 

 2,4-D 500 ppm  Mortality Ethrel 1000 ppm 1.3 100.0 

 2,4-D 100 ppm 23.2 Severe distortion of florets by assessment time Surf Excel 5.0 % 0.2 100.0 

 2,4-D 50 ppm 6.2 96.3 Surf Excel 1.0 % 1.4 100.0 

 2,4-D 10 ppm 1.2 100.0 Surf Excel 0.75 % 2.5 100.0 

 MH 250 ppm 8.5 100.0 Surf Excel 0.25 % 0 100.0 

 MH 125 ppm 2.7 100.0 Control 1.2 100.0 

 MH 75 ppm 0.3 100.0    
 

* - average of five tests starting from seventh day of spraying with an interval of 2 days. 
** - average of two tests starting from sixteenth day of spraying with an interval of 2 days. 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 

Figure A. Normal mature (top) and dehiscing anthers 

(bottom).  
 

 

for 2, 4-D at 50 ppm and GA at 150 ppm. The slight 
deviation in pollen germination in the treatments with 2, 4-
D at 50 ppm and GA at 150 ppm, maybe due to the 
natural causes which were elaborately reviewed by 
Stanley and Linskens (1974); Bots and Mariani (2005). 
Observations on the pollen germination strengthen the 
findings that the gametocides tried were ineffective in 
causing pollen sterility either by inducing sterility or 
suppressing the germination. Salgare (2004) reported 
that cent percent pollen germinability of Cyamopsis 
tetragonoloba was suppressed by all the concentrations 

 
 

 

of maleic hydrazide above 3000 µg/ml. This supports that 
concentration of maleic hydrazide used in the experiment 
may be well below the toxic levels for pollen germination 
and repeat spraying of maleic hydrazide should also be 
assessed for pollen germination. But, pollen germination 
alone is not sufficient for normal formation of zygote 
because further growth of tube in the style leading to 
fertilization should take place. Whether these chemicals 
have any role on growth and fertilization of coriander 
needs to be further evaluated.  

Though none of the chemicals had induced pollen 
sterility or suppressed pollen germination, Maleic Hydra-
zide at 125 and 250 ppm caused severe suppression of 
anther dehiscence. The chemical also suppressed anther 
protrusion and prevented the petal opening. A series of 
effects were observed in the florets under the influence of 
the maleic hydrazide over time. One of the primary 
effects was pollen agglutination. Pollen agglutination was 

first observed on 7
th

 day to 9
th

 day after spraying but a 
clear phase of agglutination started from 10 days after 
spraying. Poor opening of florets and prevention of 

protrusion of anthers were observed during 10
th

 to 22
nd

 
day. During this phase, severe agglutination of the pollen 
was observed (Figures) which caused prevention of 
pollen dispersal (suppression of anther dehiscence). The 

effect relapsed on 23
rd

 day after spraying. This phenol-
menon was further verified using another test variety 
Swathi. The effect of maleic hydrazide on Swathi followed 
the similar trend as with the case of Sadhana (Table 4).  

As early as 1969, Tadahiko Hirose reported that 
sodium 2, 2-dichloropropionate caused suppression of 
anther dehiscence in bell pepper which started about two 
weeks after treatment. However, there is a paucity of 
information on effect of gametocides on anther dehi-
scence of crop plants. Rustagi and Mohan Ram (1971) 
reported functional male sterility with the use of Mendok 
(sodium 2, 3-dichloroisobutyrate) and Dalapon (sodium 2, 

  



5 

 

  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
Figure B. Suppression of anther dehiscence with the use of  
Maleic Hydrazide 125 ppm.  

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
Figure C. Pollen agglutination in anthers affected by Maleic 

Hydrazide.  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
Figure D. Pistil of Normal floret (Left) and Maleic Hydrazide  
affected floret (Right). 



6 

 

  
 
 

 
Table 4. Maleic Hydrazide influence on anthesis and anther dehiscence.  

 
Days after spraying Observations on varieties Sadhana and Swathi 

6
th

 day to 9
th

 day Normal anthesis. Normal pollen dehiscence 

a. 10
th

 day-22
nd

 day in Sadhana Poor opening of petals. Anthers do not protrude. 

b.   10
th

 day-21
st

 day in Swathi Pollen agglutination and suppression of dehiscence. 

a. 23
rd

 day in Sadhana Relapsed to normal anthesis 

b. 22
nd

 day in Swathi  
 

 
Table 5. Percentage of fruit set in maleic hyrazide assisted crossing.  

 
 Cross combination Percentage fruit set (%) Cross combination Percentage fruit set (%) 

 Sudha x LCC-121 35.0 Sadhana x LCC-121 46.0 

 Sudha x LCC-139 42.0 Sadhana x LCC-139 38.9 

 Sudha x LCC-143 55.1 Sadhana x LCC-143 56.1 

 Sudha x LCC-149 35.3 Sadhana x LCC-149 38.9 

 Sudha x LCC-173 41.4 Sadhana x LCC-173 32.1 

 Sudha x LCC-163 42.2 Sadhana x LCC-163 70.5 

 Sudha x LCC- 215 43.1 Sadhana x LCC-215 58.5 

 Mean 42.0 Mean 48.7 
 Standard deviation 6.7 Standard deviation 13.5 

 

 

2-dichloropropionate) at 250, 500 and 1000 ppm in 
linseed where functional male sterility resulted from lack 
of anthesis, fusion and non- dehiscence of fertile anthers 
and agglutination of pollen. The results of the present 
investigation are similar to the Rustagi and Mohan Ram’s 
report. 
 

 

Use of Maleic Hydrazide in chemical emasculation of 

coriander 
 
In the treatments with repeated spraying of maleic 
hydrazide, the observations recorded through stereo mi-
croscope revealed that the persistence of severe pollen 
agglutination continued from the first day of anthesis to 
cessation of flowering in all the treatments. This is signi-
ficant as the entire flowering phase could be brought 
under the influence of maleic hydrazide thus forcing 
pollen agglutination which prevented selfing. Bagging of 
the umbels on treated plants did not result any fruit set 
further confirming the microscopic observations. In con-
trast, selfing in untreated female parents was 8.0% in 
Sudha and to 16.1% in Sadhana indicating that, without 
assisted pollination, no fruit set takes place in the umbels 
of treated plants due to prevention of anther dehiscence. 
 

 

Successful crosses with the use of Maleic Hydrazide 

 

Fourteen cross combinations were attempted involving 

two female parents and seven male parents using the 

maleic hydrazide as chemical emasculation agent which 

 

 

resulted in 42% fruit set involving with female parent 
Sudha and 48.7% fruit set involving female parent 
Sadhana demonstrating the utility of the technology 
(Table 5). The success in obtaining seed of desired 
parentage through maleic hydrazide assisted crossing 
revealed that the huge amount of crossed material can be 
generated in coriander through the use of maleic 
hydrazide as chemical emasculation agent with relative 
ease. 
 

 

Summary of the findings 
 
The present investigation identifies maleic hydrazide as 
potential male gametocide and an effective alternative for 
cumbersome hand emasculation in coriander. Spraying of 
maleic hydrazide 100 ppm from 25 DAS on wards until 
the cessation of flowering is an effective alternative to 
cumbersome emasculation in coriander and for quick 
generation of large quantity of breeding material. 

 
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Radboud Universiteit Nijmegen, Netherlands. pp: 14-28.  
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Chauhan SVS. , Vandana Singh (2002). Detergent induced male 

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Diederichsen A (1996) Promoting the conservation and use of 

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