Table Of ContentMutations, In Vitro and Molecular Techniques for Environmentally
Sustainable Crop Improvement
Mutations, In Vitro and Molecular
Techniques for Environmentally
Sustainable Crop Improvement
Edited by
M. Maluszynski
Joint FAO/IAEA Division 0/ Nuclear Techniques in Food and Agriculture,
Vienna, Austria
and
K.J. Kasha
University 0/ Guelph,
Department 0/ Plant Agriculture,
Guelph, Canada
....
"
SPRINGER-SCIENCE+BUSINESS MEDIA, B.V.
A C.I.P. Catalogue record for this book is available from the Library of Congress.
ISBN 978-90-481-6012-9 ISBN 978-94-015-9996-2 (eBook)
DOI 10.1007/978-94-015-9996-2
Selected papers have been published in Euphytica, Vol. 120, No. 3,2001
Printed on acid-free paper
All Rights Reserved
©2002 Springer Science+Business Media Dordrecht
Originally published by Kluwer Academic Publishers in 2002
Softcover reprint of the hardcover 1s t edition 2002
No part of this work may be reproduced, stored in a retrieval system, or transmitted
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EDITORIAL
The views expressed in the papers, nomenclature used and way of presentation remain
the responsibility ofthe authors concemed, who are also responsible for any reproduction
of copyright material. In certain cases only a short communication is presented. All
published papers were reviewed.
The excellent assistance of Mrs. Katayon Allaf and Kathleen Weindl, Joint FAOIIAEA
Division, Plant Breeding and Genetics Section for editorial help, retyping some
manuscripts and making corrections in English, is highly appreciated.
Contents
Preface ......................................................................................................... . . ... IX
* Mutation genetics ofsalt tolerance in barley: an assessment ofGolden Promise and
other semi-dwarf mutants .. ..................... ........................ . ........ 1
B.P. Farster
* Mapping genes for flowering time and frost tolerance in cereals using precise
genetic stocks ................................................... '" ......................................................... 17
J. W Snape, R. Sarma, S.A. Quarrie, G. Galiba and J. Sutka
* Characterization of wheat -Thinapyrum partial amphiploids for resistance to barley
yellow dwarfvirus .......................................................................................................... 27
G. Fedak, Q. Chen, A. Cameau and CA. St.-Pierre
* In vitra techniques for genomic alteration in rice plants ....................................................... 35
T Kinashita and K. Mari
* An improved in vitra technique for isolated microspore culture ofbarley ............................ .45
K.J. Kasha, E. Simian, R. ara, Q.A. Yaa, TC Hu and A.R. Carlsan
* Heterosis in crop mutant crosses and production of high yielding lines using doubled
haploid system.. .. ............. .......... .. ... ............. ... . .................................................... 55
M Maluszynski, I. Szarejka, P. Barriga and A. Balcerzyk
*Haploidy breeding and mutagenesis for drought tolerance in wheat ..................................... 75
A.J. Khan, S. Hassan, M Tariq and T Khan
* Anther culture in connection with induced mutations for rice improvement ......................... 83
Q.F. Chen, CL. Wang, YM Lu, M Shen, R. AJza, MV. Duren and H. Brunner
Production of salt tolerant rice mutants using gamma rays and anther culture ...................... 93
W Navarra Alvarez
* Induction of photoperiod sensitive genetic male steriles for use in hybrid rice seed
production .................................................................................................................... 97
J.N. Rutger
* Aluminum tolerance in triticale, wheat and rye ................................................................. 10 1
B. Y Kim, A. C Baier, D.J. Samers and J.P. GustaJsan
Evaluation of' Anahuac' wheat mutant lines for aluminum tolerance ................................. 113
A. Tulmann Neta, CE. de Oliveira Camarga, J. Lapes de Castra and
A. w.P. Ferreira Filha
*Chapters indicated with an asterisk are reprinted from Euphytica, Volume 120, No. 3 (2001).
viii
* Barley mutants with increased tolerance to aluminum toxicity .......................................... 121
M Nawrat, 1. Szarejko and M Maluszynski
* Anthocyanin mutations improving tomato and pep per tolerance to adverse c1imatic
conditions.......... .............. ............ .................. .............. .. ....... 139
B. Atanassova, S. Daskalov, L. Shtereva and E. Balatcheva
The use of somac1onal variation and in vitro selection for improvement of plants to
environmental stresses ............................................................................................... 151
J. Bouharmont, H. Batoko, P. Bertin, C Costa de Macedo, S. Lutts, N. Skali Senhaji,
V. Van Sint and G. Zhu
In vitra techniques for the selection ofBasmati rice mutants better adapted to Saline
environments. . . . .. . .................................. . . ............... 161
A.A. Cheema, MY. Saleem and MA. Awan
Rice germplasm enhancement by induced mutations in Chile. . ............................... 169
JR. Alvarado Aguila and R.P. Madariaga Burrows
Induced mutations as a method of obtaining iron toxicity resistant and high quality
rice cultivars... ......... ............ .. ...... ........ . ............................................ . . .... 175
R.E. Bacha, S. Yokoyama and T Ishiy
Improvement oftartary buckwheat by induced mutations with 60CO gamma rays ............... 183
Y. Tang, Z.X Bin and Z. Gang
Evaluation ofbarley mutants for drought tolerance: a physiology - breeding approach ...... 189
MI. Cagirgan, J. Gorham, C Toker, B. Uzun, K. Visser
* Mutations affecting nodulation in grain legumes ................................................................ 20 1
CR. Bhatia, K. Nichterlein and M Maluszynski
Summary ......................................................................................................................... 227
Index ................................................................................................................................ 241
IX
Preface
During the last thirty years, most increases in agricultural production were achieved through
high input agrieulture in areas with fertile soils and sufficient water. Intensive methods of
production with high levels of nitrogen fertilizer and pesticides were often accompanied by
environmental degradation and in some instances by pollution of the food supply. However,
rapid population growth has also led to increasing use of marginal lands, where adverse soil
and climatic eonditions are serious constraints to food production. These areas are even more
sensitive to ecological destabilization.
Environmentally sound systems of food production and land use are essential for meeting
the food security needs of developing countries. To do this, greater genetic variability is
needed within the best crop genotypes available for the areas in need coupled with better
management praetices and crop rotations. These requirements can only be realized if suitable
crop varieties are bred. These should be varieties with a much shorter growing period, suitable
for rotation, increased tolerance or resistance to diseases and pests as weil as to drought and
salinity and other adverse soil and climatic conditions.
Induced mutation techniques and other biotechnological approaches are major tools for
creating variability for stress resistance or tolerance and for manipulating and evaluating them
in efficient ways for improved germplasm. There are a number of them to ehoose from for
producing new and useful variation witrun partieular species and for particular agronomie
situations. Additionally, scientists have become intensely interested in mutations as a means to
widen and deepen our understanding of gene function. Among the many potential
biotechnologies, in vitra culture and molecular methods seem most appropriate for developing
countries ..
The transfer of technology to developing countries needs more than just produets of
biotechnology but the technology itself There are several reasons for trus. First, the best target
for improvement is local germplasm, whieh is already weil adapted and has the specific traits
acceptable to the consumers and thus can be seleeted by the local breeders. Second, it provides
local employment and enhances loeal research capability, enabling breeders to also work on
minor crops that otherwise would be ignored. However, this requires strong loeal government
involvement and support which may not be optimal and hence the need for catalytic external
assistance.
This publication contains the results of an F AOIIAEA Research Co-ordinated Projeet
(CRP) entitled "Radiation Induced Mutations and Other Advaneed Teehnologies for the
Production of Crop Mutants Suitable for Environmentally Sustainable Agriculture". These are
supplemented by information related to the topic on approaches to folIowand methodologies
available with emphasis on applications in developing countries. The publication deals only
with seed crops, mostly cereals but also tropicallegumes. Nevertheless the strategies used and
conclusions reaehed are valid for many other types of crops.
Arising from the diversity of research objectives and strategies the outcomes of the
project, although broad, are fundamentally twofold. First, a number of biotechnologies were
developed and validated in association with the use of mutants to understand the genetics of
speeific plant traits; and second, a number of mutants were developed with benefieial
characteristics and which have significant long term potential to contribute to sustainable
agriculture. For example, within the first category, the use of advanced technologies to
characterise specific mutated genes e.g. ari-e.GP responsible for semi-dwarfness plus salinity
and drought tolerance in barley, and the development of applieable in vitra selection and
x
haploid methods, means that the impact of the project will continue to yield benefits as the
understanding of a particular gene function flows through to plant improvement programmes in
many countries. Also, in vitra methods of doubled haploid production and the genetic
information resulting from this project open new possibilities for large-scale production of
radiation-induced doubled haploid mutants and high yielding doubled haploids from heterotic
mutant crosses ofbarley and other cereals.
Within the second category, was the creation of radiation-induced mutants of rice,
barley, wheat, tomato and buckwheat with increased tolerance to abiotic stress e.g. salinity,
aluminium toxicity, cold tolerance, has direct relevance to crop improvement in Brazil,
Bulgaria, Chile, China and Pakistan. Agriculture in many countries will also benefit from the
development of disease resistant varieties as a result oftechnologies applied in this project. For
example, the use of gamma radiation to generate asymmetrie in vitra fusion lead to rice lines
with resistance to blast disease, and the transfer of genes for resistance to wheat streak mosaic
virus and barley yellow dwarf virus from wild relatives into crop cultivars.
James D. Dargie
Director, Joint FAO/lAEA Division
ofNuclear Techniques in Food and Agriculture
Vienna, Austria
Mutation genetics of salt tolerance in barley:
an assessment of Golden Promise and other
semi-dwarf mutants
B.P. Forster
Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, Scotland, UK
Key words: barley, semi-dwarfmutation, ari-e.GP, salt tolerance
Summary
A review of research at the Scottish Crop Research Institute (SCRl) on the
effects of semi-dwarfing genes on salt tolerance in barley is given. Work
began in 1993 with the fortuitous and unexpected result that the cultivar
'Golden Promise' showed considerable tolerance to salt. Golden Promise is
gamma-ray induced semi-dwarf mutant of the cultivar 'Maythorpe'. The
parent and mutant cultivars are presumed to be isogenic, but show significant
differences in their responses to salt stress. The positive and pleiotropic
effects of the mutant gene, commonly known as GPert were found to be
effective in a number of genetic backgrounds. Earlier, in 1991 Frackowiak
showed that the GPert mutation was allelic to the ari-e mutants in barley. The
ari-e mutants were salt tested and found to show the same positive responses
to salt stress as Golden Promise. This supported the allelism tests, and
consequently the GPert symbol was changed to ari-e. GP. The semi-dwarf
mutant sdw.l (also known as denso) and the erectoides semi-dwarf mutant,
ert_!?2 were also tested for their effects on tolerance to salt, but did not show
any positive effects. Salt tolerance was therefore not a general phenomenon of
semi-dwarf stature but specific to mutations at the Ari-e locus in these lines.
Genetic markers (RAPDs, AFLPs and SSRs) have been used for
fingerprinting, genetic mapping, and QTL analysis. The markers have helped:
I) confirm the isogenic relationship between Maythorpe and Golden Promise,
2) clarify the confusion over the pedigree of Golden Promise, and 3)
genetically map the ari-e.GP locus and examine its pleiotropic effects.
M. Maluszynski and K.l. Kasha (eds.),
Mutations, In I1tro and Molecular Teclmiquesfor Environmentally Sustainable Crop Improvement, 1-16.
© 20021AEA.
2
Background
Early work on salt tolerance at The Scottish Crop Research Institute (SCRI) was aimed at
screening variation in cultivated (Horde um vulgare) and wild (H. spontaneum) barley lines.
There was a particular emphasis in screening lines which had been used in developing doubled
haploid lines (DHs) as a segregating DH population could be exploited in genetic mapping of
quantitative trait loci (QTLs) for salt tolerance. Various barley lines were subjected to a
hydroponics experiment of two treatments, 0 and 175 mol m-3 NaCI, and salt tolerance
measured in terms of shoot sodium content (mg NaJg dry weight, Forster et al., 1994). From
this initial screen a breeding li ne HS26/55C was found to be the most salt tolerant spring
barley. HS26/55C is an interesting line in that it was bred as areplacement for the popular, but
mildew susceptible cultivar 'Golden Promise'. lt was developed from a cross between cv.
Golden Promise and a wild barley (H. spontaneum HS26) in an attempt to transfer rnildew
resistance from the wild species into Golden Promise (Thomas et al., 1988). lt was therefore
presumed (wrongly) that the salt tolerance ofHS26/55C had been inherited from the wild line
HS26 along with mildew resistance, as it was known that certain wild barley populations were
weB adapted to saline environments. HS26/55C was therefore tested along with its mildew
resistant donor, HS26 and its recurrent parent, Golden Promise. After 4 weeks exposure to
175 mol m-3 NaCI the shoot sodium contents ofHS26/55C and Golden Promise were found to
be similar; 23.6 and 21.8 mg NaJg dry weight respectively (no significant difference). However,
HS26 had more than twice the amount of shoot sodium (54.7, P<O.OOI, Forster et al., 1994).
lt was conc1uded that the salt tolerance of HS26/55C was inherited from Golden Promise,
further work on Golden Promise was initiated.
The Golden Promise story
The GPerl mutation was induced by gamma-ray irradiation ofthe cultivar 'Maythorpe' in 1956
(Fig. I). The aim was to induce shorter stiffer straw in a good malting quality barley by
mutation. The result was the production ofthe highly successful malting quality barley, Golden
Promise, a direct mutant of Maythorpe (Miln Marster, described in Sigurbjornsson and Micke,
1969), GPert stood for Golden Promise erectoides.
Golden Pro mise offered barley growers a unique combination of highly desirable
agronomic characters such as earliness, short stiff straw, easy combining ability, good
resistance to grain and ear loss and to grain splitting with good malting quality. Golden
Promise was particularly suited to agriculture of Northern Britain. In 1967 it was inc1uded on
the recommended list of cereals for England, a year later it was recommended for Scotland
where it dominated Scottish barley acreage from the 1970s to the mid 1980s. Golden Promise
was susceptible to powdery mildew (Erysiphe graminis sp. hordei), but the widespread use of
effective chemicals allowed Golden Promise to maintain its dominant position and take an
increasing share of the UK market. The introduction of Golden Promise to Scotland
revolutionised cereal growing to the benefit of the farmer, malt ster, brewer and distiller
(fungicide manufacturers also profited), and enabled ScotIand to co me to the fore as a
producer of malting barIey. Golden Promise became a standard for malt quality and is still
grown today under contract for specialist distillers. The introduction of Golden Promise not
only presented farmers with a new plant model (semi-dwarf stature), but also provided an
additional morphological gene source for breeding. Golden Promise contains the recessive
