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Euphytica

DOI 10.1007/s10681-015-1385-4

Inheritance of high iron and zinc concentration in selected


bean varieties
F. Mukamuhirwa • G. Tusiime • M. C. Mukankusi

Received: 5 January 2014 / Accepted: 30 January 2015


Ó Springer Science+Business Media Dordrecht 2015

Abstract Micronutrient malnutrition especially iron non-additive gene effects are important in determining
(Fe) and zinc (Zn) has disastrous consequences for the the expression of high Zn and Fe concentration even
more vulnerable members of the human society, though additive gene effects contributed more. Nar-
especially poor women and pre-school children in row sense heritability of Fe and Zn was high estimated
developing countries. The common bean (Phaseolus at 71 % for Fe and 83 % for Zn. Significant maternal
vulgaris L.), an important grain legume directly and reciprocal effects suggest also that cytoplasmic
consumed by humans provides cheap and important inheritance is involved in Zn and Fe concentration.
quantities of protein and calories and is an excellent Strong positive correlation between Fe and Zn
source of some minerals (Fe and Zn) and vitamins concentrations (r = 0.75) was observed suggesting
which qualify it as perfect food. Unfortunately, many that these micronutrients are not independently inher-
of the adapted bean market class varieties are low in ited. The positive relationship between Fe and Zn plus
these nutrients. Therefore there is need for biofortifi- the negative relationship observed between Zn and
cation of these varieties with Fe and Zn through plant seed size (r = 56) suggest that Zn and Fe accumula-
breeding and genetic engineering in order to con- tion is controlled multigenically or oligogenically with
tribute to improved health of bean consumers. The some genes affecting the concentration of both
objective of this study was to determine the mode of minerals. The variability in seed mineral concentra-
inheritance of high Fe and Zn concentration in selected tion among crosses was larger for Fe (47–77 ppm)
bean varieties. Six parents high and low in Fe and Zn than for Zn (28–38 ppm).
were crossed following a full diallel crossing design.
Populations were advanced from F1 to F2. F3 seed were Keywords Gene effects  General and specific
analyzed for Fe and Zn concentration using X-ray combining ability  Heritability
fluorescence. The study showed that both additive and

F. Mukamuhirwa (&)  G. Tusiime


Introduction
Department of Agricultural Production, Makerere
University, P. O. Box 7062, Kampala, Uganda
e-mail: fmukamuhirwa@yahoo.cm Increasing the amount of Fe and Zn concentration in
common bean grain may contribute significantly to
M. C. Mukankusi
improving the health status of individuals dependent
CIAT Africa, National Agricultural Research
Laboratories- Kawanda, P. O. Box 7065, Kampala, on beans as a staple food (House et al. 2002).
Uganda Significant increases in the amount of bioavailable

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Zn and Fe in beans can be made through traditional Materials and methods


plant breeding techniques (Nchimbi-Msolla and
Muhamba 2010) based on the existing wide variation Study location and plant materials
of Fe and Zn concentration among bean genotypes.
Therefore the knowledge of inheritance is critical in The study was carried out in the screen house at CIAT
designing appropriate breeding strategies for incorpo- Uganda based at National Agricultural Research
rating a particular trait into economically useful Laboratories (NARL) in Kawanda, Uganda from June
populations. The aim of this study was to contribute 2011 to August 2012. Six bean inbred lines were used
to the development of high Fe and Zn beans through in this study, they included, three high and three low
studying the mechanism of inheritance of iron and Fe and Zn concentration bean varieties based on
zinc. The objectives of this study were: HarvestPlus data. The high Fe lines included;
KAB06F2827- developed at Kabete-CIAT breeding
1. To study the gene action governing Fe and Zn
program, NUA99 which is among several lines
concentration in beans;
deployed in Africa from CIAT-Colombia as high Fe
2. To determine the combining ability among six
bush Andean beans, RWR 2154 a released variety in
common bean varieties for Fe and Zn
Rwanda developed by the Rwanda Agriculture Board
concentration.
(RAB) bean breeding program. The low Fe parents
3. To estimate the role of maternal effects control-
included, CAL96 a red mottled bean developed at
ling Fe and Zn concentration in beans;
CIAT and released in Uganda. It is highly susceptible
4. To estimate narrow sense heritability (h2) for Fe
to all major diseases and is used as a local disease and
and Zn concentration in common bean
high yield check and is low in Fe. DOR500 is a CIAT
populations;
line currently used as a universal low Fe check.
5. To estimate heterosis on Fe and Zn concentration
RWR2076 is a RAB bred line used as a local yield
in the F2 generation of common bean crosses;
check in Rwanda.
6. To estimate the magnitude of relationship be-
Characteristics of these entries are outlined in the
tween iron and zinc concentration in F2
figure below (Fig. 1).
populations.

NUA 99 RWR 2076 RWR 2154

KAB06F2.8-27 DOR 500 CAL 96

Fig. 1 Seed of parents used in the study of inheritance of high iron and zinc concentration

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Population development generation. The remaining seed was bulked into four
replicates for iron and zinc analysis at Rubona
The study germplasm was grown and crossed follow- Agriculture Research Station, Rwanda for XRF
ing a full diallel design for testing maternal and analysis (Paltridge et al. 2012). XRF uses X-rays to
paternal effects (Hallauer and Miranda in Tadess et al. energize atoms within sample, causing them to
2008; Jones 1965; Crusio 1987). Planting of the fluoresce. Energy and intensity of fluorescence pro-
crossing block was done on a mixture of forest black vides information about elemental make-up. The
soil, lake sand and decomposed farm yard manure in a machine scans each sample for 100 s with spinning
ratio of 3:1:1 (currently used by CIAT). A carefully of sample cup (use 4 g of grain (7 ml) to analyze Fe
mixed composite sample was packed in new paper and Zn and records intensities of emitted X-rays.
bags, labelled twice and analyzed at Kawanda Agri- (Pfeiffer and McClafferty 2007; Harvest Plus 2011).
cultural Research Laboratories (KARL). Soil analysis Inheritance and gene action of Fe and Zn was studied
was done following the protocol of Soils and Soil based on results obtained from the analysis of F3 seeds.
Fertility Management Programme of National Agri-
cultural Research Laboratory, actually at Kawanda. Data analysis
Soil was analyzed for iron (Fe), zinc (Zn), acidity
(pH), organic matter (OM), nitrogen (N), Phosphorus Analysis of variance was performed by the ANOVA
(P), Potassium (K), Calcium (Ca), and Magnesium procedure of Genstat 14th Edition (VSN International
(Mg). The experimental soil had medium to very high Ltd., 2012). Significant treatment effects were separat-
nutrients. Apart from iron (140 ppm) that was very ed using the least significant difference (LSD).
high and sufficient in concentration, calcium Correlation and regression analysis were carried out
(1,720 ppm), magnesium (360 ppm), potassium with linear regression analysis of Microsoft Excel.
(425 ppm) and zinc (5 ppm) concentrations were very
high but not sufficient. Phosphorus (17 ppm) concen- Estimation of general and specific combining
tration was high and not sufficient. Organic matter ability
(3.7 %), pH 5.1 and nitrogen (0.21 %) were medium
but not sufficient. Each parent per cross combination General and specific combining ability were deter-
was planted in five buckets and repeated weekly for mined following Model 1 of Griffing’s analysis
4 weeks. Four crossing blocks were set up. N.P.K 17: (Griffing 1956). General combining ability (GCA)
16: 16 fertilizer was added to experimental soils on a for male and female parents as well as the specific
weekly basis from germination to pod formation. combining ability (SCA) effect means were obtained
Thirty grams of N.P.K was dissolved in 10 l of water, following the formula developed by Hallauer and
from which 50 mls of the fertilizer solution was Miranda in Tadesse et al. (2008). GCAi = mean of
applied to each pot weekly. The experiment was crosses of parent ‘‘I’’- grand mean, while SCAij = -
watered in the morning; although in some cases, cross mean - (grand mean ? GCAf ? GCAm).
watering was done in the afternoon when deemed
necessary. The crossings were done in 6 9 6 full Estimation of heritability
diallel after emasculating the female parents. The bulk
method was used to advance F1 to F2 and F2 to F3. Narrow sense heritability (NSH) was estimated from
variance components. Broad sense heritability, was
Evaluation of developed populations for Fe and Zn calculated as H = VG/VP, and narrow sense herit-
concentration ability, h2 = VA/VP where H and h2 are broad sense
heritability and narrow sense heritability respectively,
At F3, seed was harvested by individual plant and put VA is additive variance, VG is genotypic variance and
in clean new paper envelopes to avoid contamination VP is phenotypic variance (Hill et al. 2008). Baker’s
with dust and dirt. These were hand threshed under ratios were used to predict perfomance of crosses
conditions that kept the seed as free of dirt and dust as based on GCA values. The closer the ratio is to 1 the
much as possible. Thereafter, five seeds per plant were greater the chances of predicting performance based
randomly selected for advancement to the next on GCA (Baker 1978). Table 1 below summarizes the

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Table 1 Formulae used for estimating heritability and Baker’s ratio


Formulae

Baker’s ratio 2*re2GCA/(2*re2GCA ? SCA re2)


NS-CGD (genotype mean basis) 2*re2GCA/(2*re2 GCA ? re2 SCA ? re2)
BS-CGD (genotype mean basis) (2*re2GCA ? re2 SCA)/(2*re2 GCA ? re2 SCA ? re2)
NS-CGD (narrow sense coefficient of genetic determination) = estimate of narrow sense heritability (h2) BS-CGD (broad sense
coefficient of genetic determination) = estimate of broad sense heritability (H)

formulae used for heritability and Baker’s ratio based concentration) parent and WP is the mean of worse
on GCA and SCA values (Griffing 1956; Baker 1978). (low Fe/Zn concentration) parent.
When for a given trait the narrow sense heritability
is high, the trait is highly heritable and when is small it
is lowly heritable. High heritability estimates indicate Results
that additive gene action is more important for that
trait and selective breeding of the best to the best Gene action determining iron and zinc
should produce more desirable progeny. Low esti- concentration
mates on the other hand indicate that probably non-
additive gene action such as overdominance, dom- Twenty seven successful crosses out of the planned 30
inance and epistasis is important. If heritability is high, were evaluated. The analysis of variance showed
individual selection is recommended and if heritability significant difference among crosses (P \ 0.001) for
is low, family selection is recommended (Behera both Fe and Zn concentration. General combining
2007). Narrow sense heritability was also estimated by ability effects were highly significant at P \ 0.001 for
mid-parent offspring regression analysis. The regres- both Fe and Zn concentration while the specific
sion coefficient ‘‘b’’ is an estimate of narrow sense combining ability effects were also significant at
heritability (Falconer and Mackay 1996). P \ 0.001 and P \ 0.01 for Fe and Zn concentration,
respectively. GCA effects accounted for 64 and 82 %
for Fe and Zn, respectively, while SCA accounted for
Estimation of heterosis effects for iron and zinc 36 and 18 % for Fe and Zn, respectively. This indicated
concentration observed at the F2 generation that additive gene action was more important than
non-additive gene action implying that the highest Fe
In this study, heterosis was determined for the F2 and Zn concentration progeny may be produced by
populations that involved six parents (KAB06F2.8-27, crossing the two parents with high GCA effects.
NUA 99, RWR 2076, RWR 2154, CAL 96 and DOR Reciprocal effects were also highly significant
500) high and low in Fe and Zn concentration. The (P \ 0.001) for all traits evaluated (Table 2). Highly
amount of heterosis was calculated by comparing the significant maternal and non-maternal reciprocal ef-
mean of F2 progeny over mid-parental value, over fects were observed for both Fe and Zn concentration
better parental value and over the worse parental value indicating the importance of cytoplasmic 9 nuclear
in respect of Fe and Zn concentration using the gene interaction effects in accumulating high Fe and
formula of Falconer and Mackay (1996). Mid-parent Zn concentration.
heterosis was calculated as: MPH = (F2 - MP)/ The variability in seed mineral concentration among
MP * 100 where F2 is the mean performance of the crosses was larger for Fe (47–77 ppm) than for Zn
F2 population and MP is the mean of two parents. (28–38 ppm). Transgressive segregation for higher Fe
Similarly, heterobeltiosis was obtained as the differ- and Zn was observed where some crosses (RWR
ence in the mean performance of the mean of F2 to 2076 9 KAB06F2.8-27 and RWR2076 9 DOR500
their high or low iron/zinc parent, that is; BPH (better (one direction) showed higher Fe or Zn concentration
parent heterosis) = (F2 - BP)/BP * 100; WPH than the higher parent (Table 3). The crosses of
(worse parent heterosis) = (F2 - WP)/WP * 100 RWR2076 9 KAB06F2.8-27 and RWR2076 9
where BP is the mean of the better (high Fe/Zn DOR500 performed well for both iron and zinc since

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Table 2 Mean squares for Source of variation DF Iron (ppm) Zinc (ppm) 100 SW Seeds per plant
different traits evaluated
along iron and zinc Entry (Tot.) 32 46.23 7.273 384.44 61.77
concentration in crosses of
GCA 5 171.01*** 32.40*** 2,167.52*** 319.66***
bean varieties high and low
in seed iron and zinc SCA 15 19.64*** 1.73** 72.14*** 12.85***
concentration Reciprocals 12 27.48*** 3.73*** 31.85*** 19.02***
Crosses 27 23.13*** 2.62*** 54.24*** 15.71***
ns, **, *** = not Maternal effects 6 19.59** 4.86*** 51.82*** 28.93**
significant, significant at Non maternal effects 6 35.37*** 2.61** 11.88ns 10.53**
1 % and 1 % level Error 32 4.34 0.56 7.17 2.29
respectively

their parents showed good general combining ability concentration. RWR 2076, KAB06F2.8-27 and DOR
suggesting additive genetic effects (Fig. 2). 500 had positive GCA effects suggesting that these
varieties contributed to high Fe and Zn concentration
Estimation of combining ability effects in the crosses that involved them. CAL 96, NUA 99
for developed crosses and RWR 2154 had negative GCA effects suggesting
that they contributed to low Fe and Zn concentration in
General combining ability the crosses that involved them.
Differences in GCA values (Fig. 2) are attributed to
Positive GCA effects were desirable in this study additive gene effects. Positive GCA effects of DOR
because they indicated the bean line’s contribution to 500 and high Fe concentration in the crosses that
high Fe and Zn concentration while the negative GCA involved it yet it is a universal low Fe check suggests
effects represent contribution towards low Fe and Zn that it might be having recessive genes for high Fe and

Table 3 Means of F3 seeds for iron and zinc concentration along their parents in ppm
Female parents Male parents
CAL96 NUA99 RWR2076 RWR2154 KAB06F2.8-27 DOR500

Iron concentration CAL960 46.79 58.29 54.29 54.29 65.29

NUA99 52.29 55.79 59.04 50.79 63.54 62.54


RWR2076 60.29 53.29 64.04 70.04 76.79

RWR2154 51.79 62.79 66.79 53.79 58.54

KAB06F2.8-27 58.29 63.29 75.79 65.04 66.04 68.29


DOR500 58.65 57.29 60.04 60.29 64.29 57.29
Zinc concentration CAL96 27.99 30.57 30.57 31.57 34.49

NUA99 30.57 32.24 33.99 27.57 32.24 35.74


RWR2076 31.99 31.99 35.24 34.74 37.99

RWR2154 28.57 32.99 30.74 31.24 34.24

KAB06F2.8-27 31.07 36.57 37.74 31.99 34.24 37.49


DOR500 34.70 33.99 35.74 34.74 34.49 35.24
Figures in the diagonal are parents while figures in bold are crosses with distinctly transgressive segregation with good performance
for Fe and Zn respectively

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impacting on high Fe concentration in these crosses.


SCA effects were positive for CAL 96 9 RWR 2154,
NUA 99 9 KAB06F2.8-27, RWR 2076 9 KAB06
F2.8-27, CAL 96 9 DOR 500, RWR 2076 9 DOR
500, RWR 2154 9 DOR 500 indicating the presence of
non-additive gene effects governing high Zn concen-
tration in these crosses. Differences in SCA are
attributed to non-additive effects including dominance
and epistasis gene effects (Table 4).

Estimation of the role of maternal effects on Fe


Fig. 2 Parental GCA effects of F3 seeds of crosses for high iron and Zn concentration in beans
and zinc concentration. Average SE GCA was 0.40 for iron
concentration and 0.15 for zinc concentration Reciprocal crosses RWR 2076 9 CAL 96, RWR
2154 9 NUA 99, KAB06F2.8-27 9 RWR 2076,
Zn concentration. Similarly NUA 99and RWR 2154 had positive reciprocal effects in F3 generation
contributed to low Fe concentration yet they are high (Table 5) implying that iron concentration was only
Fe parents suggesting epistasis and or overdominance higher when RWR 2076, RWR2154 and KAB06F2.8-
gene effects. 27 were maternal parents in these crosses.
Crosses CAL 96 9 NUA 99, CAL 96 9 DOR 500,
Specific combining ability NUA 99 9 RWR 2076, NUA 99 9 DOR 500, RWR
2154 9 DOR 500 had high iron concentration when
SCA effects were positive for CAL 96 9 NUA 99, CAL CAL 96, NUA 99 and RWR 2154 were maternal
96 9 DOR 500, RWR 2076 9 RWR 2154, RWR parents and lower Fe concentration when NUA 99,
2076 9 KAB06F2.8-27 and RWR 2076 9 DOR 500 DOR 500, RWR 2076 were maternal parents in these
indicating the presence of non- additive gene effects crosses. On the other hand reciprocal crosses RWR
Table 4 SCA effects in F3 seed of crosses between beans high in iron and zinc concentration in ppm
Trait Parent Genotypes Males parents
CAL96 NUA99 RWR2076 RWR2154 KAB06F2.8- DOR500
27

Iron Female CAL96 -3.86 2.37 -1.60 -0.58 -2.10 4.73


parents NUA99 0.59 -5.00 0.89 0.75 0.40
RWR2076 -3.08 4.93 4.28 2.93
RWR2154 -2.81 1.67 -0.80

KAB06F2.8- -4.11 -0.70


27
DOR500 -6.55
AVG SE SCA 1.27
Zinc Female CAL96 -0.90 0.13 -0.65 0.51 -0.94 1.37
parents NUA99 0.25 -0.49 -0.83 0.85 0.09
RWR2076 0.27 -1.86 1.19 0.60
RWR2154 1.01 -0.69 0.59

KAB06F2.8- -0.88 -0.35


27
DOR500 -2.32
AVG SE SCA 0.47

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Table 5 Reciprocal (maternal and non-maternal) effect values in crosses between beans high and low in iron and zinc concentration
Trait Parent Genotypes Male parents
CAL96 NUA99 RWR2076 RWR2154 KAB06F2.8-27 DOR500

Iron Female parents CAL96


NUA99 -3.00
RWR2076 ?3.00 -2.88
RWR2154 -1.25 ?6.00

KAB06F2.8-27 -0.12 ?2.88

DOR500 -3.32 -2.63 -8.38 ?0.88 -2.00


AVG SE RECIP 1.41
Zinc Female parents CAL96
NUA99 0.00
RWR2076 ?0.71 -1.00
RWR2154 -1.50 ?2.71

KAB06F2.8-27 ?2.16 ?1.50

DOR500 ?0.11 -0.87 -1.13 ?0.25 -1.50


AVG SE RECIP 0.53
?,2 : positive and negative reciprocal effects indicating the importance of cytoplasmic 9 nuclear gene interaction effects in
accumulating high Fe and Zn concentration

2076 9 CAL 96, RWR 2154 9 NUA 99, KAB06 genotype mean basis. The h2 was estimated at 0.83 and
F2.8-27 9 NUA 99 and KAB06F2.8-27 9 RWR 0.71 for zinc and iron respectively suggesting that Fe
2076 had high Zn concentration when RWR 2076, and Zn concentration can be improved more rapidly
RWR 2154 and KAB06F2.8-27 are female parents and with less evaluations. High Baker’s ratio of 0.90 and
low Zn concentration when CAL 96, NUA 99, and 0.78 for zinc and iron respectively were observed
RWR 2076 are maternal parents in these crosses. (Table 6).
Crosses involving CAL 96 9 RWR2154, NUA Narrow sense heritability was also estimated by
99 9 RWR 2076, NUA 99 9 DOR 500, RWAR mid-parent offspring regression analysis where the
2076 9 DOR 500 and KAB06F2.8-27 9 DOR 500 regression coefficient b represents narrow sense
had high zinc concentration when CAL 96, NUA 99, heritability (h2). The mid-parent offspring regression
NUA 99, RWR 2076 and KAB06F2.8-27 are maternal analysis was also significant (P \ 0.001) with the
parents and low Zn concentration when RWR2154, adjusted regression coefficient b of 1.05 and 1.17 for
RWR 2076 and DOR 500 are maternal parents in these iron and zinc, respectively. This was an indicator of a
crosses. The reciprocal effects are also explained by very high narrow sense heritability suggesting that
the maternal and non-maternal effects generated by genetic effects on iron and zinc concentration were
diallel analysis. These indicated that the cytoplasm of very high. Therefore high iron and zinc concentration
these varieties contributed to high Fe and Zn concen- in the F2 plants (F3 seed) generation can be reliably
tration of the crosses involving these parents. predicted based on parental performance. F2 plants (but
F3 seed) data indicated that 46 and 55 % of the total
Estimation of narrow sense heritability (h2) for Fe variation in iron and zinc concentration respectively of
and Zn concentration in 27 common bean F3 seed populations was accounted for by the parental
populations iron and zinc concentration (Fig. 2). A relatively high
r2 indicates high predictability of crosses from parents
The narrow sense heritability (h2) was estimated by since the mid parent has relationship with the cross.
narrow sense coefficient of genetic determination at This is shown by the scatter plots below (Fig. 3).

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Table 6 Baker’s ratio and heritability estimates for Fe, Zn, 100 SW and seed per plant
Component Source of variation Iron Zinc 100 SW Seeds per plant

Variance components GCA 15.43 2.95 200.03 28.34


SCA 8.50 0.65 36.09 5.65
RECIPS 11.57 1.59 12.34 8.36
Heritability Baker’s ratio 0.78 0.90 0.92 0.91
NS-CGD (genotype mean basis) 0.71 0.83 0.90 0.88
BS-CGD (genotype mean basis) 0.90 0.92 0.98 0.96

Heterosis and heterobeltiosis for iron and zinc concentration F2 crosses had positive heterobeltiosis
concentration observed at the F2 plants generation to high Fe parent. On the other hand, 12 out of 14
high 9 low Fe concentration F2 crosses had positive
Positive heterosis is desirable as it indicates the heterobeltiosis to low Fe parent while 13 out of 13
superiority of the F2 to either mid-parent, high Fe/Zn low 9 high Fe concentration crosses had positive
concentration parent and low Fe/Zn concentration heterobeltiosis to low Fe parent.
parent. The heterosis was used to understand the The negative heterobeltiosis observed for the
contribution of different parents to Fe/Zn concentra- crosses involving NUA 99 as high Fe parent may
tion and may help in the selection of desirable crosses suggest the involvement of over-dominance effects in
(those with high positive heterosis) in a breeding these crosses.
scheme. Thirteen out of 14 high 9 low iron concen- The positive heterosis observed with crosses with
tration F2 crosses had positive relative/mid-parent KAB06F2.8-27 indicated that this bean line may be a
heterosis while 9 out of 13 low 9 high Fe concentra- good source of Fe concentration since it still resulted
tion F2 crosses had positive relative/mid-parent in a better offspring. Crosses involving KAB06F2.8-
heterosis. The positive relative heterosis varied from 27, RWR 2076 resulted into high Fe concentration
1.33 to 26.58. High 9 low Fe crosses had higher offsprings when they were female parents than when
heterosis level compared to low 9 high crosses indi- they were male parents, indicating possession of
cating that high Fe concentration levels were obtained cytoplasmic genes that contribute to Fe concentration.
when high Fe parents were used as mothers than vice Crosses involving DOR 500 (a low universal check) as
versa. Crosses with positive heterosis show the male parent resulted into high Fe concentration
presence of joint action of a favorable combination offspring suggesting recessive genes for high Fe.
of genes at different loci. Six out of 14 high 9 low Fe Even though complete dominance effects was ob-
concentration F2 crosses had positive heterobeltiosis served for some crosses (CAL 96 9 RWR2154, DOR
to high Fe parent while 8 out of 13 low 9 high Fe 500 9 CAL 96, DOR 500 9 NUA 99, KAB06F2.8-

Fig. 3 Regression of iron/


zinc concentration, F2
plants progeny (F3 seed)
iron/zinc concentration on
mid-parent for iron/zinc
concentration for 27
populations

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27 9 RWR2154 and RWR 2154 9 DOR 500), the Fe concentration (r = 0.75; P B 0.001. The relationship
level of these crosses was low. Most of crosses between Zn and 100 seed weight (r = -56;
involving CAL 96, NUA 99, RWR 2154 had negative P B 0.001) was unfortunately significant but negative.
heterosis suggesting that they contributed to low Zn The relationship between number of seeds per plant
concentration in these crosses. Interestingly, crosses and Zn concentration was also significant (r = 0.50;
involving parents KAB06F2.8-27and RWR2076 had P B 0.01 (Table 8).
high positive heterosis for zinc as they had for iron.
Though high heterosis and high Zn concentration were
obtained by using each of the parents as a female, the Discussions
use of KAB06F2.8-27 as a female parent was more
beneficial than any other parent indicating that this The variability in seed mineral concentration among
line possesses cytoplasmic genes that contribute to crosses was larger for iron (47–77 ppm) than for zinc
high Zn concentration. The positive heterosis ob- (28–38 ppm) with significant and strong positive
served with crosses with KAB06F2.8-27 indicated that correlations between iron and zinc (r = 0.75) and a
this bean line could also be a good source high Zn strong negative correlation between zinc and seed size
concentration. Even if RWR 2076 9 DOR 500 had (r = 0.56). Phenotypic variability in seed iron and
the same high parental Zn concentration, high hetero- zinc concentration observed in this study depended on
sis and good performance were observed when RWR the genotypes and environmental conditions under
2076 was the female parent and DOR 500 the male which the study was carried out. Similar observations
parent. Eight out of 14 high 9 low Zn concentration were earlier reported by Cichy et al. (2009).
F2 crosses had positive relative heterosis while 10 out Both general combining ability (GCA) and specific
of 13 low 9 high Zn concentration F2 crosses had combining ability (SCA) were found to be important
positive relative heterosis. The positive relative in determining progeny performance, indicating that
heterosis varied from 0.00 to 10.01. Low 9 High Zn both additive and non-additive effects were important.
crosses had higher heterosis level compared to Baker’s ratio (1978) for the relative importance of
high 9 low crosses (Table 7). Two out of 14 9 low GCA and SCA variance components was 0.90 for zinc
Zn concentration F2 crosses had positive heterobel- and 0.78 for iron, demonstrating that additive gene
tiosis to high Zn parent while 6 out of 13 low 9 high effects were more important than non-additive ones in
Zn concentration F2 crosses had positive heterobel- determining the expression of high zinc and iron
tiosis to high Fe parent. On the other hand 12 out of 14 concentration even though SCA effects were also
high 9 low Zn concentration F2 crosses had positive highly significant.
heterobeltiosis to low Zn parent while 12 out of 13 Data in this study support Hill et al. (2008) in their
low 9 high Zn concentration crosses had positive evidence from empirical studies of genetic variance
heterobeltiosis to low Zn parent. The negative components found that additive variance typically
heterobeltiosis observed in high 9 low Zn concentra- counts for over half and often close to 100 % of the
tion crosses and low 9 high Zn concentration crosses total genetic variance. The results also support the
as well as the negative heterosis and heterobeltiosis to study of Blair et al. (2009) where they found that
low Zn concentration observed in the crosses involv- inheritance of higher iron and zinc concentration was
ing NUA 99 as high parent may suggest the involve- mainly additive. Mostly additive gene action suggests
ment of over-dominance gene effects in these crosses. that selection in early segregating generations should
be effective for improving these traits. The use of non
Estimation of the magnitude of relationship destructive measurements for Fe and Zn such Marker
between iron and zinc concentration in F2 assisted selection is recommended. Based on these
populations research findings one should first select genotypes
with high zinc concentration and evaluate them for
Simple correlations were calculated among mineral high iron concentration and then discard those with
mean values to examine the relationship between the low iron since zinc is correlated to iron with higher
concentrations of the two minerals. Significant and heritability. Narrow sense heritability of iron and zinc
positive correlations were found between Fe and Zn was high according to Johnson et al. (1955) and was

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Table 7 Midi-parent heterosis and heterobeltiosis for zinc concentration observed on F2 plants (high Zn 9 low Zn) and their
reciprocals (low Zn 9 high Zn) crosses
Cross code Mean Zn concentration of parents F2’s Zn Mid-parent Mid-parent Heterobeltiosis (%)
concentration Zn heterosis
High Zn Low Zn High Zn Low Zn

NUA99 9 CAL96 32.24 27.99 30.57 30.11 1.51 -5.18 9.21


CAL96 9 NUA99 32.24 27.99 30.57 30.11 1.51 -5.18 9.21
RWR2076 9 CAL96 35.24 27.99 31.99 31.61 1.19 -9.22 14.29
CAL96 9 RWR2076 35.24 27.99 30.57 31.61 -3.31 -13.26 9.21
RWR2154 9 CAL96 31.24 27.99 28.57 29.61 -3.53 -8.55 2.07
CAL96 9 RWR2154 31.24 27.99 31.57 29.61 6.60 1.05 12.79
KAB06F2.8-27 9 CAL96 34.24 27.99 31.07 31.11 -0.15 -9.26 11.00
DOR500 9 CAL96 35.24 27.99 34.70 31.61 9.76 -1.53 23.98
CAL96 9 DOR500 35.24 27.99 34.49 31.61 9.09 -2.13 23.22
RWR2076 9 NUA99 35.24 32.24 31.99 33.74 -5.19 -9.22 -0.78
NUA99 9 RWR2076 35.24 32.24 33.99 33.74 0.74 -3.55 5.43
RWR2154 9 NUA99 32.24 31.24 32.99 31.74 3.94 2.33 5.60
NUA99 9 RWR2154 32.24 31.24 27.57 31.74 -13.14 -14.49 -11.75
KAB06F2.8-27 9 NUA99 34.24 32.24 36.57 33.24 10.01 6.80 13.43
NUA99 9 KAB06F2.8-27 34.24 32.24 32.24 33.24 -3.01 -5.84 0.00
DOR500 9 NUA99 35.24 32.24 33.99 33.74 0.74 -3.55 5.43
NUA99 9 DOR500 35.24 32.24 35.74 33.74 5.93 1.42 10.86
RWR2154 9 RWR2076 35.24 31.24 30.74 33.24 -7.52 -12.77 -1.60
KAB06F2.8-27 9 RWR2076 35.24 34.24 37.74 34.74 8.64 7.09 10.22
RWR2076 9 KAB06F2.8-27 35.24 34.24 34.74 34.74 0.00 -1.42 1.46
DOR500 9 RWR2076 35.24 35.24 35.74 35.24 1.42 1.42 1.42
RWR2076 9 DOR500 35.24 35.24 37.99 35.24 7.80 7.80 7.80
KAB06F2.8-27 9 RWR2154 34.24 31.24 31.99 32.74 -2.29 -6.57 2.40
DOR500 9 RWR2154 35.24 31.24 34.74 33.24 4.51 -1.42 11.20
RWR2154 9 DOR500 35.24 31.24 34.24 33.24 3.01 -2.84 9.60
DOR500 XKAB06F2.8-27 35.24 34.24 34.49 34.74 -0.72 -2.13 0.73
KAB06F2.8-27 9 DOR500 35.24 34.24 37.49 34.74 7.92 6.39 9.49

Table 8 Relationship of iron and zinc concentration along with seed size and number of seeds per plant (based on entry means)
Trait Fe Zn 100 SW Seeds per plant

Fe 0.75*** -0.12NS 0.04NS


Zn -0.56*** 0.50**
100 SW –0.82***
Seeds per plant
**, *** = significant at P B 0.01 and 0.001 respectively. Fe = Iron, Zn = Zinc, 100 SW = 100 seed weight

estimated at 71 % for iron and 83 % for zinc. The interactions are involved in zinc and iron concentra-
significant maternal and reciprocal effects suggest also tion. A maternal effect was also found for seed protein
that cytoplasmic inheritance and cytoplasmic-genic in beans (Noubissié et al. 2012). Maternal effects play

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an important role at most developmental stage parents KAB06F2.8-27 and RWR2076 should be used
(Variath et al. 2009) and this may have a direct impact to maximize opportunities for improvement of iron
on the selection process and the progression of and zinc and they should be used as female parents.
segregating population in genetic improvement pro- The parent DOR 500, even though is a low parent
grams. The diallel analysis revealed that additive should be used as male parent because it should be
effects as well as non-additive genetic effects influ- having recessive genes for high Fe and Zn concentra-
ence iron and zinc concentrations. Therefore the tion. F2 populations such as KAB06F2.8-27 9 RWR
progeny performance on iron and zinc may not be 2076, and RWR 2076 9 DOR 500 performed better
fully predictable based on parent performance alone. with significant heterosis for both iron and zinc
Selection of parents with high concentrations of Fe concentration. Significant heterosis was also observed
and Zn, along with screening crosses in early in cotton by Maria Khan Panni et al. (2012) and in
generations, especially for zinc (Baker’s ratio of 0.90 wheat by Khatun et al. (2010). Therefore crosses
compared to 0.78 for iron) should be effective in involving KAB06F2.8-27 and RWR2076 as maternal
improving the concentrations of these minerals in bean parents and DOR 500 as paternal parent performed
seed. Correlation in iron and zinc concentrations well for iron and zinc and should be selected and
(r = 0.75) observed in this study support the results of advanced to enhance the iron and zinc concentration in
Blair et al. (2009) which showed co-localization of these crosses.
QTL responsible for high zinc and iron in beans. This
may be interpreted as evidence that accumulation of Acknowledgments We are very grateful to Makerere
University, AGRA, CIAT, Harvest plus project and RAB for
both minerals are of similar inheritance. Cichy et al.
the support to this work.
(2009) also reported correlations of Fe and Zn up to
0.53 and the co-localization of QTL for iron and zinc.
The co-localization of QTL for seed iron and zinc
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