and Methods

Plamt Breeding: Principles

50

some species, the degree orincompati-

of
In
End-of-Season Pollination. season in mature
bility is reduced towards the
end of the fioweringthe usefulness of this
controversi l reports on
But there are
plants.
technique.
In some species, e-8Trifolfum, Lycopersicon,
High Temperature. exposure of to temperatures upto
pistils
Brassica, Oenothera etc.
60C indutes pesudo-fertility.
5ystem, .e.g., in
Irradiation. In the single-1ocus gametophytic
or gamma-rays induces a
irradiation with X-rays
Solanaceae, acuteself-incompatibility.
temporary loss of
Grafting. Grafting of a branch onto aoother branch of theofsame
self-
to reduce the degree
plant or of another plant reported
is
incompatibility in Tryolíum pratense. There is only one report
and the machanism of this reduction is not
on this phenomenon,
known.

Double Pollination. In some species,self-incompatible matingss

become possible when incompatible pollen is applied as a mixture

with a compatible pollen,

or it is applied after pollination with aa

compatibl: pollen.
number of otber techniques have been tried with
Other Techniques.of. are not commonly used. These
varying degrees success, but they
techniques are treatment of flowers with carbon monooxide,
injecting styles With immunosuppressants, application of electrical
potential difference of about 100 V between the stigma and pollen
grains, treatment of pistil with phytohormones and with protein
synthesis inhibitors, and steel brush pollination.

MALE STERILITY

Male sterility is characterised by nonfunctional pollen graBns,

whilefamale gametes function normally. toceurs in nature
sporadically, perhaps due to mutation. Male sterility is classified into
chree groups: (1) genetic, (2) cytoplasmic, and (3) cytoplasmic
genetic.

Genetic Male Sterility

Genetic male sterility is ordinarily governed by a siogle reces-
sive gene, ms, but dominant genes governing male sterility are
also known, eg, Male sterility alleles arise
in safiower.
spontaneously or may be artificially induced. A male sterile line
may be maintained by crossing it with heterozygous male
fertile
plants.Such a mating produces 1:l male sterile and male fertile
plants (Fig. 3.6).
Utilization in Plant Breeding ienetic male sterility may be used in
hybrid seed production. The progeny from ms msX Ms ms crosses
are used as female, and are
interplanted with a homozvgous male
Modes of Reproduction and Pollination Control 61

fertile (Ms Ms) pollinator. Thc genotypes of the ms nms and Ms ms

lines are identical except for the ms locus, i.e., They are
are known as male sterile (A) and maintainer (B) lines, isogenic, und
rcspectivcly.
The female line would, therefore, contain both male sterile and male
fertile plants; the latter must be identified and removed before
pollen shedding. This is done by identifying the male fertile plants in
seedling stage either due lo the pleiotropic ellect of the ms gene or
due to the phenotypic effect of a closely-linked genc. Pollen dispersal
from the male (pollinator) ine should be good for a satisfactory
Seed set in th: fem ileline. However, generally pollen dispersal is
poor and good, closely-linked markers are rare, Roguing of male
fertileplants from the female line is costly as a result of which the
cOst of hybrid seed is higher. Duc to these dificulties, genetic
male sterility has been exploited commercially only in a few countries.
In USA, it is being successfully used in castor. In India, it is being
Used lor hiy brid sced production of arhar (C. caian) by some private
seed companics, e.g: Maharashtra Hybrid Sted Co. Ltd., India,
produced and sold 50 Q seed of a hybrid variety of arhar. Suggestions
have been made for its use in several other crops, e.g.. cotton,
barley. tomato, sunflower, cucurbits etc., but it is not yet practically
feasible.

Inberitance of Male St erlty

Parents ms ms Ms Ms
(Malestcrile), (Male fertile)
Fi Ms
(Male fertile)

I Ms Ms, 2 AMs ms, 1 ms ms

Ratio 3:
Male fertile Male sterile
Maintenance of A Male Sterile I.ine
Mulc sterilc strain A Male fertile Crossthe
themale
male seriie line
strain B
with fertilc sib
ine
IS ms X Ms Ms

s ms* Ms M 3 The Fi (Ms ms) is back.

Male sterile (line A), Male fertile crossed with the ma l
ne
3 1 Ms mis: ms nis ntalned by sib mating.
Malc tcrile x Male steril Sced from the male sterile
(Sccd nut harvested) (Seca harves tcd) plants only 1s haryested.

31Msms: 1 ms ms
Male
Maintaincd by sib mating
fertilex Male sSterile sced from the male sterile
(Sced not harvested) (Seed harvested) p'ants only is harvested.

Maintained indcioitely

Fig. 3.6. Inberitancc genetic male strarility. Male sterility

of gene designated as ms; the dominant allele Ms pro- is produced by a
recessive nuclear
duces male fertility. During the maintenance of male stevile lines,
sibmatin8 is achieved through natural pollination.
 52 Plamt Breeding : Princlples and Methods
Cytoplasmie Male Sterility
This type of male sterility is determined by the
3.7). Since
cytoplasm (Fig.
the cytoylasm of a zygote comes primarlly from cg8 cell,
ne progeny of such male sterile plants would always be male sterile.
Cytoplasmic male sterility is known in many plant species, some of
which are crop plants (Table 3.2). Cytoplasmic male sterility may
D transrerred easily to a given strain by using that strain as a polli.
nator recurrent paren) in the successive generations of a backcross
programme (Fig. 3.8). After 6-7 backcrosses, the nuclear genotype of
the male sterile line would be almost identical to that of the recurrent
pollinator strain. The male sterile line is maintained by crossing it

CYTOPLAsM STERILE
NUCLEAA GENE NONRESTORER
TIS uriABLETO cOUNTERACT
THE
EFFECT OF THE STERILEE
CYTOPLASM
MALE STEAILE

CYTOPLASM FERTILE
NUCLEAR GENE NON RESTORER

MALE FERTILE

- MALE STERILE MALE FEATLE MALE STERILE

Fig 3.7.
Cytoplasmic male sterility. Male sterility is caused by the cytoplasm
(depicted by S). The F cyloplasm is the normal cytoplasm. Ir a
restorer gene (the dominant allele of r) becomes available, the male
sterility will be included ia the
of male sterility shows strict cy
cytoplasnic genetic group. Tais typo
toplasmic inheritance.

with the
pollinator strain used as the recurrent
parent inwith
thethat
back-
Cross programme since its nuclear genotype is identical of
the male sterile line. Such a
male maintain
tainer line or B line as it is used tofertile line the
is knowD as
the main
male sterile line.
The male slerile lIne is also known
as the A line. There is
ble evidence that the
gene or genes conditioning cytoplasmic considerTa
sterility, particularly in maize, reside in mitochondria, and may male
be
located in a plasmid like clement.
 Modes of Reproduction and Pollination Control
63

Transfor of Male Sterllity to a New Strain

STRAIN AA
STRAIN B

Cross the male

Ierlile strain B to
a citoplkismic male
stetile strain A

MALE STERILE MALE FERATILE

(i) F would be
mole sterile

MALE STERILE
A

OSTRAIN B
(ii)
(ii) Backcross the
ostrain B
50%, of the
nuclent genes
would be from
strain B

(0 Backcross to

BC

MALE STERILE
( STRAINB
(ii)
Strainof
75%
Duclear
the
8enes
would he irom
Strain B.

REPEATED BACK CROSSES

() This is the male

sterile version
of strain B

BCe Or BC
(>99% nuclear
genes from 5.
(ii) Maintained by
maungwith
male ertile
strain B.
MALE STERILE STRAIN
B

Fig. 3.8. Transfer of the male sterile ceytoplasm from strain A to strainB.
Strain A is used as the non-recurrent parent in the backerosspro
will bo
&ramme with stran B. After 6-7 backcrosses, the progeny
similar to strain B their
in oucleargenotype. The straia B serves as
the maiotaíner of the Dew male sterile strain.
 Methods
Plant Breeding : Principles and
64
ma le sterility io some

Table 3.2. Cytoplasmic and cytoplasmic-genetic

important crop plants.

Restorer genes Remerks

(nucleus) Cytoplasm
Crop sperie
cms-C (2. mays)" Onc (RS)* Spontaneous e
Maize, (Zea mays) version relatively
high

cms-S {Z. mays)* One (RSa)** Commercially

used

cms-T (Z. mays)* Two (Rf1, Rf1)** Most commonly

used, slightly re
tards growth and
yield (2-4%)
susceptibility to
Helminihosporlum
Icaf blight

Poor growth,
Nicotiana tahacum Nicotinna debneyl floral abnormali-
Nirotiana megalosíphon ties, conimercially
Nicotiana bigelcvli
not USCd.

Triticum aestivum Triticum ti1nophrevii Two (RS1. RS:) Commercially

Aegilops cauiata rom 7. iinio not used.
pheevif

Triticum durum Aegilops o vata Comniercially

not used

Gossypium hirsutum Gossypium anomalum Commercially

Gossypium arboreumm not used

Sorghum bicolor Milo (S. bcolor) One (Msc) Commercially

used

Pennisetum americanum Tifton (P. anericanum)" . Commercially

Ludhiana (P. americanum) used

Helianthus anuus H. annuus Two (Rf1, Rfi) Commercially

USea

Oryza sativa Wild rice (WA type) Eporophytic Widely used

O. sativaF spontanea actioon used
Chinsurah Boro I Gametophytic used
action
(BT type)
Gambiaca (Gam type) used
O-Shan-Tao-Bai used

Brossiea juncea 3.napus Two geDes proposed to

cumulativo be used
efect

Mutant cytoplasm.
Restoration of fertility
Iertility ot pollen grains by
Rfi and
depends on Rfa
showssporophytic
the genotype
control,
of the plant,
thatthat
while
is,
by RSsshows gametophytic control, f.e., fertility of pollen grains depends
on their owa genotYpes.
 65
and Pollnaton Control
Modes of Reproduction
male sterility may be utiliz-
Ueilization inPlant Brceding. Cytoplasmic in
seed in certain ornamental species, or
ed for producing hybrid of economic value.
But in those
species
where
a vegetalive part is no use because
crop plants
where seed economic part, it is of
is the
would be male sterile.
the bybrid progeny

Cytoplasmic-Genetic Male Sterility

nuclear
This is a
case of cytoplasmic male sterility where a

male sterile line is known. The

gene for restoring tertility in the is tound in certain strains
fertility restorer gene, R, is dominant and
of the species, or may be transterred fronm a ielated species, e.g., 1n
restores male fertility in the male sterile
wheat (Tableit 3.2). This gene The cases obf
line, hence 1s known as restorer gene (Fig. 3.9).
cytoplasmic male sterifity would be included in the cytoplasmic-
restorer genes for them would be dis-
genetic system as and when
covered. It is likely that a restorer gene would be found for all the
cases of cytoplasmic male sterility ifa thorough search were made
rice and
This system is known in maize, Jowar, bajra, sundower,
wheat (Table 3.2).
The plants would be male sterile in the presence of male
sterile cytoplasm if the nuclear genotype wererr, but would be male
fertile if the nucleus were Rr or RR (Fig. 3.9). New male sterile Iines
may be developcd following the same procedure as in the case of
cytoplasmic system (Fig. 3.8). But the nuclear genotype of the polli
nator strain uscd in the transter must b e rr, other wise The Tertility
would be restored. The development of new restorer strains is some
what indirect (Fig. 3.10). First a restorer strain (say R) is crossed with
(A). male
athemale sterile line The resulting fertile plants are used
female parent in repeated backcrosses with the strain (C), uscd
as
as
the recurrent parent, to which the transfer of restorer gene is desired.
In eachgeneration, male sterile plants are discarded, and the male
fertile plants are used as females for backcrossing to the strain
CThis acts as a selcction device for the restorer gene R during
1he backcross
progranime. At the end of the backcross programme,
a restorer line isogenic to the strain C would be recovered.

Utilization in Plani Breeding. The cytoplasmic-genetic male sterility

1S used commercially to produce hybird seed in maize, bajra and
jowar. A generalised scheme for producing a double cross in maize
1S
presented in Fig.3.11. Alternatively, two fertility restoring inbreds
may be used to produce the single cross If. One of tbe two inbreds,
inthis case, will
bave to be detasselled for use
as the female parent.
Thus all plants in tke double will
cross be male fertile, since the
single cross II will be homozygous for the restorer gene R. For
producing single crosses, the scheme tor production of the
cross II as presented in Fig. 3.I1 is followed. Single
A triple cross may be
produced by crossing single cross 1 (Fig: 3.11) with a fertility restor
ing inbred so that all the plants in the triple cross would be male
fertile,
 56 Plant Breeding: Principles and Methods

Cytoplasm sterile
(O Nirclear gene
resorter cene nonrestorer, e , recessive allele of the

MALE STERILE

( MALE FERTLE
Cytoplasm fertile (nonsterile)
NOCIear gEDe nonrestorcr

Cytoplamgene
sterile
restorer in homozygous (RR) or
Nuclear
heterozygous (Rr) state
Effect of the sterile cytoplasm negated by the
restorer gene

( BOTH MALE
FERTILE
Various Genotypes and Phenotypes

-
MALE STERILE
MALE FERTILE
-( MALE STERILE

(O (R RF (
MALE FERTILE
MALE FERTLE
MALE STERLE

- MALE FERTi
I MALE FERTILE 1MALE STER".E
AALE STERILE

Results of Varlous Matingssterility is due to a sterile

Fig. 3.9. Cytoplasmjc-genetic Ismale
sterility. Male which is usually
cytoplasm;1ertility restored by a tberestorer gene, indicates that the
a dominant ouclear gene. FS in cytoplasm
fertile (F) or sierile (S).
cytoplasm may be eitber
 Modes of Reproduction and Pollination Control 67

Restorer line crossed to

cytoplasmic male sterile

RR ine.This allows selec tion

for R gene in the segregat
ing generations.
CSTOnEa

The resalting male fertile

YAR ( 1S
C
CrosSed to the strain
to wDich R 1S to
transferred The F, IS used
as temale to retain the
De

VALtPATA male sterile cytoplasm.

STAA "

Male fertile progeny

back-crossed to strain C.
Strain C is used as thee
recurrent male parent.
DCARcED

EA

MALE STEANE
O Male fertile progeny is
back-crossed to strain C
Strain C is used as
male
(DNSCARCE
-76BAC oss

SE #TH

AADH
( Male fertile progeny self-
pollinatedd

Male fertile progeny selfed.

EAn
Individua! plant progenies
grown in tbe next
.e
MALE ST
genera
tion and nonsegregating
tDsCA1OE
progenies selected.
PROSENY TEST NONSEGnEGA.G PoGENES

Fig.3 10. Transfer of restorer gene R from a restorer strain

Dew strain (strain (strain R) to a
C
Origin of Male Sterile Cytoplasm. Male sterile
spontaneously in nature or may be produced by the cytoplasm
breeder. arises
The
various sources of the male sterile
cytoplasm are given below.
 68 Plant Breedlng: Princtples und Methods

MALE STERLEJ

SINGLE
CROSS 1A B)
9 MALL STENILE)

NOREO
tNONASTOER-
MLE FERT)

GROSS

UH MALLILNTT
STLAILIT Y
NOREDC MIC
YALE SIEALED
SINOLL
(MALL F L t t i
NBRED D
OHER
ALePERTLE)

Fig. 3.11. A generalised scneme for the production of a double cross in maize
using cytoplasmic-gcnctic male sterility.

L. Spontaneous Mutatlon. Mutant male sterile cytoplasms arise

spontaneously in low frequencies. Mutant cytoplasms have been
isolated in maize, bajra and sunilower (Table 2.2). It is likely that
an extensive search would lead to the isolation of male sterile cyto
plasms in almost every crop species.
2. Interspecific Hybridization. Transter of the full somatic chromo
some complement of a crop species, through repeated backcrossing,
into the cytoplasm of a related.wild species often leads to cytoplasmic
male sterility. Male sterile cytoplams of wheat have been derived
from Triticum timopheevii or Aegilops caudata. Other examples are
listed ia Table 2.2. In cross-pollinated crop species, the male sterile
cytoplasms have generally originated through mutation, while in
self-pollinated crops they have been transferred from related spccies.

3. Induction through Ethidium Bromide. Ethidium bromide is a

potent mutagen for cytoplasmic genes or plasmagenes. Male sterile
Cytoplasm may be induced by seed ireatment with ethidium bromide.
Such mutants have been induced in some plant species, e.g., Petunia.

LAmitations of Cytoplasuic-Genetic Male Sterilty for Use in Plant

Breedlng. There are several problems in the utilization of cytoplasmic-
genetic male sterility in hybrid seed production in many species.
These problems are listed below.

1. Undesirable Efects of the Cytoplasm. Male sterile cytoplasms

generally have undersirable side etfectsTable 2.2). For example,
the Texas cytoplasm (cms-T) in maize, by far the most successful
cytoplasm commercially, slightly retards growth, yield (2.4%),
Modes of Reproductian and Polination Control
59
plant neight and leaf number;
pollen shedding: and induces earlier silking and
Helminthosporium leaf
makes the plants highly delayed
extreme sensitivity blight. This susceptibility issusceptible
of _mitochondria due to theoto
toxin produced from cms-T genotypes to a
side cffects is by therare. fungus. A g0od cytoplasm without any
indeed The
could not be used due to their male-sterile cytoplasmssidein tobaccoo
Restorer genes only restore severe undersirable
remove the side eflects of male fertility: they are eflects.
the male sterile unable to
2.
Unsatisfactory cytoplasms.
of fertility is notFertility Restoration. In many cases,
be used in the satisfactory. As a result, these restoration
production of hybrid seed. sources cannot
3.
Unsatisfactory Pollination. Natural
satisfactory, except in w.nd-pollinatedpollination
crops
is
often not
reduces the
production like maize.
cost. Io some species,
of
hybrid seed, and thereby increasesThis its
of male e.g. Capsicum, this has
sterility
would always be
in prevented
hybrid seed production. Poor the use
In rice, this problemmajor
a
pollination
problem in self-pollinators, e.8.,
in the is
morning, generallyovercome by regular tripping ofwhcat. ears
4.
using rope.
a
Modifier genes may reduce the
male sterility. During backcrosses, effectiveness of cytoplasmic
sterile cytoplasms, the while transferring the male
disturbed. This nuclear genetic
may lead to some polenbackground may also be
sterile lines. production by the unale
5.
Sometimes,
in cytoplasm may' also be
contributed by the sperm
which, the long run, may lead to a breakdown of
sterility mechanismn. the male
6. Male sterility mechanisms may break down partiaBly under
certain environmental
duction by the maleconditions resulting in some pollen pro
encountered in maize, bajrasterile lines. This problem has beena
and
7. jowar.
In crops Iike wbeat, polyploid
nature of the crop and undesira
ble linkages with the
a suitable
restorer
restorer (R) line.
gene make it very difficult to
develop
SUMMARY
sexual and asexuai reproduction Osexually, Some crops
reproduciag Cr pS show both
foerpollinated, sel.
Opening of
openiog of iowers alter polination, clustering of
Cross-pollination is anthers over stigma etc.
promoted by monoecy, dioecy, dichogany:
bility ora combination of these mechanisms. sell-incompali
pollinated crops are highy heterozygous and Asexually
geDerally
reproducing and cross
exhibit sirong inbreed
1ng oepression. be sell-pollinated species are homozygous and
inbreeding depression. The do not show
the
response of crop species diierent-breeding
to methods are primarily based
inbreeding. Thus in self-pollinated specieson
&Variety consists of bomozygous
beterozygOsity bas to be retained or plants, while in cross-pollinated species
restored. Thus the modo of reproduction