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Development and evaluation of a new high yielding and better fibre quality mutant NIAB-824 of cotton through pollen irradiation.


The success of all conventional cotton breeding approaches is highly correlated with the genetic variability present within the existing germplasm. However, if the desired trait is not present or/and linked with other undesirable traits in the existing germplasm, then the cross breeding may not be worthwhile. In such cases recombination of genes is to be sought out to achieve the desired objectives. Recombination process plays a major role to induce genetic changes and the recombination of linked genes is brought about by crossing over. Generally in eukaryotic cells radiation treatments are known to enhance crossing over in proximal region adjacent to the centromere. Thus irradiation of [F.sub.1] plants especially during premeiotic stages is further known to enhance crossing over in proximal region adjacent to the centromere resulting in further enhancing the genetic variability in [F.sub.2] population. Increased variability in [F.sub.2][M.sub.2] for quantitative traits has been reported in rice (Jalil Miah and Yamaguchi, 1965). Moreover radiation as well as several chemicals are reported to increase somatic recombinations (Vig, 1973). Exposure of seed to ionizing radiation's has resulted in creating genetic variability in different crop species and many plant breeding programmes have shown the feasibility of radiation plus selection as a direct method of varietal improvement (Carnelius, 1973, Micke, et al., 1987, Iqbal et al., 1991, Iqbal et al., 1994). Irradiation of male parent pollen before cross- pollinations resulted in the induction of mutations in cotton (Pate and Duncan, 1963 and Krishnaswami and Kothandaraman, 1976). The studies carried out by Aslam and Stelly (1994), and Aslam et al., (1994) and Aslam (2002) have shown that treatment of pollen with low doses of gamma rays (5 Gy to 20 Gy) before cross-pollinations are suitable to induce useful genetic variability in cotton. The present research studies were aimed at to create genetic variability through crosses with irradiated male parent pollen, selecting the desirable recombinants from the segregating populations and their evaluation for high yield potential and wider adaptability in the cotton growing areas of Punjab for confirmation.

Materials and Methods

The well adapted and higher yielding local cotton variety NIAB-78 was crossed with an exotic line REBA -288 using irradiated male parent pollen at 10 Gy of gamma rays before cross-pollinations during the year, 1996-97. At maturity the seed cotton was collected from the bolls obtained from successful crosses and ginned to produce [M.sub.0] seed. [M.sub.1] population was grown from M0 seed at a spacing of 30 cm and 75 cm from plant to plant and row to row respectively. At maturity the seed cotton was collected from [M.sub.1] population and instead of pooling one locule per boll from all the [M.sub.1] plants together, we picked seed cotton one locule from each boll of each [M.sub.1] plant separately. The [M.sub.2] population comprising of about more than three hundred individual plants was studied and selection for the desirable mutants/recombinants was carried out. The selections from the [M.sub.2] was based upon, early maturity, better plant type, higher yield and better yield components etc., along with resistance to CLCuV disease. Of these, 12 promising mutants were grown in [M.sub.3] generation in replicated plant progeny test (RPT) with three replications. The size of the individual plot was 0.75 m x 10 m. The breeding behaviour of these progenies was studied in [M.sub.3] generation and one higher yielding progeny was selected. The same was studied in plant progeny rows in [M.sub.4] generation to confirm its higher yield potential and to see its breeding behavior/uniformity. Finally the progeny L-824 was selected from [M.sub.6] and bulked for evaluation under the name of NIAB-824. During the years i.e. 2005-2006, various trials (zonal yield trials, NCVT and DCR etc.) were conducted at NIAB, at farmer's fields etc. for confirmation of the high yield potential and wider adaptability. Moreover, various other related studies i.e. on earliness, heat tolerance, reaction towards insects and diseases particularly ClCuV disease were carried out.

Results and Discussion

The [M.sub.1] generation results revealed that the plants were faster in growth and had hybrid vigour for various traits. The [M.sub.1] generation plants showed resistance to CLCuV disease under severe natural disease epidemiology. It was noted that the [M.sub.2] generation plant progenies were generally of varied nature and some of the individual plants possessed desirable combination of certain economic traits along with resistance against CLCuV disease under high disease infestation. The boll weight and yield of the promising mutants selected from [M.sub.2] population ranged from 3.5-5.0 gram and 211-396 grams respectively. Then the mutant was evaluated in the succeeding segregating generations, the results indicated that the mutant progeny, L-824 have high yield potential. The morphological studies carried out indicated that the plant of NIAB-824 is moderately hairy, semi-compact sympodial type with fruit bearing monopodia. It has desirable leaf foliage and medium plant stature. The length of the sympodial and monopodial and internodes of the main stem are shorter than those of the parents and prevalent commercial varieties. Compared with the tall plants forming a canopy at the top, this medium plant (4-5.5 feet) is better suited for better photosynthetic activity and better yields. This plant type makes the agricultural operations such as pesticides spray, picking, inter-culture, etc., also easier. The yield performance of NIAB-824 was compared with standard varieties during 2002-2003 to 2005-06. NIAB-824 gave 5996.1 Kg/ha yield during 2002-03, higher than the standard CIM-473, 4105 Kg/ha (31.5% higher) table 1. Similarly during 2003-04 NIAB-824 gave higher yield 4315.6 Kg/ha than the standard CIM-499, 2658 Kg/ha (38.4%higher) respectively table 1. During 2004-05 CIM-499 gave 3658 Kg/ha while NIAB-824 4515.6 Kg/ha yield respectively, which is significantly 18.9% higher than the CIM-499 table 1. While in the year 2005-06 NIAB-824 surpassed two standard varieties CIM-499 and CIM-506 with yield of 4992.8 Kg/ha, 3580 Kg/ha and 2880 Kg/ha (28.28 and 43.3% higher) respectively table 1. The mutant NIAB-824 has better fibre quality traits. The morphological characters of NIAB-824 make its growing period shorter with faster squaring quality, and it matures in 150-160 days with high fruiting load to give high yield. The seeds of NIAB-824 are bold dull white and fuzzy with greenish tinge.

The cotton mutant NIAB-824 was studied in DGR trials during 2004-05,which are held on Punjab province basis under Director Cotton. The mutant NIAB-824 out of 24 cotton varieties gave yield of 2554 Kg/ha and remained at position i, iii, iv, vii, at CRSS Khanewal, CRI, Fsd, CRSS Jhang, CRS Sahiwal and PSC Khanewal respectively (Table 2). Various trials were conducted at NIAB, and on Farmers Fields and coordinated vaietal trials organized by PCCC, Karachi, and Director Cotton Research Institute, Faisalabad. The results revealed that at NIAB, on the average the NIAB -824 gave 31.5 % higher yield than CIM-473 during 2002-03 and during 2003-2004 it gave 38.4% higher yield than CIM-499 and during 2005-06 28.28% higher yield than CIM-499 and 43.3% higher than CIM-506 respectively table 1. The candidate cotton variety NIAB824 was studied in DGR trials 2005-06. NIAB-824 out yielded other cotton varieties with yield of 2022 Kg/ha, FH-207, VH-156, BH-164, MJ-6, RH-51, NIAB-884 with yield of 1998, 2003, 1568,1751,1840 and 1979 Kg/ha respectively. (Table3). The results of National Coordinated Varietal Trial (NCVT) held under PCCC, on Pakistan basis, during the year 2005-06, the mutant NIAB-824 got yield of 2521 Kg/ha, which is more than TH-84/99, 2096 GH-99, 2154 and BH-162 with yield of 2454 Kg/ha respectively. When calculated on the basis of standard plant population basis the candidate variety depicts yield of 2913 Kg/ha. (Table 4). The field response of different cotton strains against CLCuV in DGR trial at NIAB Faisalabad during 2004-05 was analyzed. NIAB-824 was having 10.45 % disease index and moderately susceptible disease reaction as compared to FH-2006 28.37 % disease index and highly susceptible disease reaction, FH-115 20.48 % disease index and highly susceptible disease reaction, NIAB-884 12.86 % disease index and moderately susceptible disease reaction. CIM-534 28.06 % disease index and highly susceptible disease reaction table 5. It depicts the better performance of mutant NIAB-824 against present threat of cotton leaf curl virus (CLCuV). The results revealed that NIAB-824 was resistance to CLCuV under high inoculation in the field. Twenty cotton varieties in NCVT (four replications) and twenty-four varieties in DGR trial (three replications) for the year 2005-06 were evaluated in field under natural inoculation of cotton leaf curl virus under normal plant protection measures. Normal agronomic practices and plant protection measures were followed to keep the crop in good conditions. Percent disease index was recorded using the rating scale.

Final observations revealed the fact that all the entries were prone to CLCuV- B infection with varying degree of responses. H-151-F2, TH-35/99, MNH-786, NIAB-824, MNH-789 in NCVT and MNH-786 in DGR were least effected while GH-99, TH-84/99 in NCVT and BH-164, SLH-284, FH-115 and N-4 in DGR were highly susceptible (Table 7). Through grafting following results were observed, ten pots each for twenty varieties in NCVT 2005-06 and twenty-four in DGR 2005-06 were sown (4-5 delinted seeds per pot) in glass house. Plants were thinned to one plant per pot two weeks after germination. These plants were graft inoculated with CLCuV-B following the bottle shoot grafting method. Results indicated that the inoculated germplasm exhibited 100% disease within 9-15 days post grafting with 6E disease severity and were rated as highly susceptible (Table 7). This situation persisted till the end of the experiment as no recovery was observed even after 120-150 days of grafting.

The results on earliness and physiological attributes of NIAB-824, showed that the flowering started earlier in NIAB-824 and is better tolerant to high temperature as compared to other varieties. Moreover it is also apparent that NIAB-824 had shorter boll maturity period thereby confirming the earliness of NIAB-824. The results of studies carried out on relative development of sucking pests and bollworms complex on different cotton varieties indicated that NIAB-824 was comparable or some time better with respect to sucking pest's complex development than other varieties. Whereas it had relatively less infestation of bollworms and due to its early maturity it can escape the peak attack period of pink bollworm and fewer number larvae enter into diapause thus reducing the carry-over source for infestation for the next year. The fibre characteristics of NIAB-824 were tested at NIAB. The results indicated that the all value have been comparable and acceptable (Table 6). NIAB-824 has G.O.T. of 37.7%, staple length of 27.0-30.0mm, fineness of 4.0-4.6[micro]g/in, strength of 94 TPPSI, fibre maturity 83-90%(Table 6). The pollen irradiation approach seem to be better than seed irradiation, since in order to create genetic variability through seed irradiation, the whole genome is to be irradiated, which ultimately disturb the whole genetic makeup of the exposed individual. Consequently the most of the changes occurring in the population are the somatic/non-heritable changes. Therefore large [M.sub.2] population may be more than 12,000 individual plants is required, to select desirable mutants (Iqbal, et al. 1994). Since the irradiated pollen is a germ cell and after fertilization only half of the genome of the developing zygote/embryo, receives the irradiation, hence the occurrence of major changes is minimized as observed in case of seed irradiation. The results confirmed that the pollen irradiation is a valuable technique, which can be employed to improve crop plants most effectively. Moreover incase of seed irradiation usually from each [M.sub.1] plant the seed cotton from each locule per boll is collected and then pooled to have [M.sub.1] seed to grow [M.sub.2] population and consequently large population has to be screened for selecting the desired genotype. But through the use of pollen irradiation technique, each [M.sub.1] plant has to be grown separately as plant progeny rows to develop [M.sub.2] population, which facilitate to carryout selection. Since most of the progenies carried micromutations/point mutation due to optimal radiation dosages applied to pollen before fertilization and no major abnormalities were noticed which may help to achieve uniformity earlier as compared to seed irradiation.

Moreover the male gamete is to be irradiated at low doses of gamma rays before fertilization and therefore more recombinations are brought about due to enhanced chaismata formation/crossing over during meiotic stages of cell divisions. The results reported above have clearly illustrated that from a very small [M.sub.2] population even less than 1000 plants, higher rate of mutations/recombinations was achieved through pollen irradiation. Therefore the results obtained clearly confirmed the earlier findings (Jalil Miah and Yamaguchi, 1965, Vig, 1973, Wang, 1990). Moreover, the method of gamete treatment was found easier to apply than that of zygote/seed treatment. Irradiation of male parent pollen before cross-pollinations resulted in the induction of mutations in cotton (Pate and Duncan, 1963 and Krishnaswami and Kothandaraman, 1976). These results are in accordance with the earlier findings [Aslam and Stelly (1994), Aslam et al., (1994) and Aslam (2000)], that the treatment of pollen with of low doses of gamma rays (5 Gy to 20 Gy) before cross-pollinations are suitable to induce useful genetic variability in cotton, G. hirsutum L.



Aslam, M. and D.M. Stelly, 1994. Attempted egg-transformation by pollen irradiation in the cotton genus, Gossypium. Bangladesh J. Nuclear Agric., 10: 1-8.

Aslam, M., R.M.S. Iqbal, M.B. Chaudhry and A.A. Bandesha, 1994. Pollen irradiation in Cotton (Gossypium hirsutum L). Pak. J. Bot., 26: 341-346.

Aslam, M., 2000. Utilization of Pollen Irradiation Technique for the Improvement of G. hirsutum L. Pak. J. Biol. Sci., 3(11): 1814-1816.

Carnelius, T.J., 1973. A new cotton variety MCU-7 by X-ray irradiation. Mutat. Breed. Newsl. 2.

Doak, C.C., 1934. A New Technique in Cotton Hybridising. Heredity, 25: 201-204.

Iqbal, R.M.S., M. B. Chaudhry, M. Aslam and A. A. Bandesha, 1991. Economic and Agricultural Impact of mutation breeding in cotton in Pakistan-a review. Plant Mutation Breeding for Crop Improvement. IAEASM-311/7, 1: 187-201.

Iqbal, R.M.S., M.B. Chaudhry, M. Aslam and A.A. Bendasha, 1994. Development of a high Yielding cotton mutant, NIAB-92 through the use of induced mutations. Pak. J. Bot., 26: 99-104.

Jalil Miah, M.A. and H. Yamaguchi, 1965. The variation of quantitative characters in the Irradiated progenies of two rice varieties and their hybrids. Radiat. Bot., 5: 187-196.

Krishnaswami, R. and R. Kothandaraman, 1976. Response of cotton pollen to gamma-Irradiation. Ind. J.Genet.Plant Breed, 36: 16-19.

Micke, A., B. Donini and M. Maluszynski, 1987. Induced mutations for crop Improvement-a Review. Trop. Agric. (Trinidad), 64: 259-278.

Pate, J.B. and E.N., Duncan, 1963. Mutations in cotton induced by gamma irradiated Pollen. Crop Sci., 3: 136-138.

Siddig, M.A., 1968. Genetics of resistance to cotton leaf curl virus in Sakil cotton. J. Agri. Sci. Camb., 70: 99-103.

Vig, B.K., 1973. Somatic crossing over in Glycine max (L) Merrill: mutagenicity of Sodium aside and lack of synergistic effect with caffeine and mitomycin C. Genetics., 75: 265-277.

Wang, L.Q., 1990. Induced Mutation for Crop Improvement -a review. Plant Mutation Breeding for Crop Improvement. IAEA-SM-311/3, 1: 9-32.

(1) M. Aslam, (2) M. Ashfaq, (1) Tahir Saeed, (2) Sami Ul Allah and (2) Muhammad Sajjad

(1) Nuclear Institute for Agriculture & Biology, P.O Box 128, Jhang Road, Faisalabad Pakistan

(2) Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad, Pakistan

M. Aslam, M. Ashfaq, Tahir Saeed, Sami Ul Allah and Muhammad Sajjad: Development and Evaluation of a New High Yielding and Better Fibre Quality Mutant NIAB-824 of Cotton Through Pollen Irradiation: Am.-Eurasian J. Sustain. Agric., 3(4): 715-720, 2009

Corresponding Author: M. Ashfaq, Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad, Pakistan

Table 1: Yield performance of NIAB-824 compared with standard
varieties during 2002-2003 to 2005-06

Mut. /Var.     CLCuV             Boll         Yield/       GOT %
               reaction          weight       ha (Kg)

2002-2003      NIAB-824 R        4.5          5996.1       38.0
CIM-473        R                 4.0          4105.0       38.5
2003-2004      NIAB-824 R        4.1          4315.6       38.2
CIM-499        R                 3.5          2658.0       38.3
2004-05        NIAB-824TB        4.1          4515.6       38.4
CIM-499        TB                3.5          3658.0       38.2
2005-06        NIAB-824 TB       4.1          4992.8       37.5
CIM-499        TB                3.5          3580.0       38.0
CIM-506        TB                3.5          2880.0       38.0

Mut. /Var.     Fibre

               length (mm)       fineness          strength
                                 [micro]g/in       (TPPSI)

2002-2003      28.5              4.4               94.0
CIM-473        28.5              4.5               93.0
2003-2004      28.6              4.3               94.5
CIM-499        28.3              4.5               93.0
2004-05        28.6              4.2               93.0
CIM-499        283               4.5               92.0
2005-06        28.5              4.1               92.0
CIM-499        28.0              4.5               --
CIM-506        29.0              4.4               --

% Increase of NIAB-824 over
CIM-473 = 31.5

% Increase of NIAB-824 over
CIM-499 = 38.4

% Increase of NIAB-824 over
CIM-499 = 18.9

% Increase of NIAB-824 over
CIM-499 = 28.28
CIM-506 = 43.3

Table 2: Yield performance of NIAB-824 in DGR trials (2004-2005)

Code    Variety      Mean         Code     Variety      Mean

V1      FH-2006      2021         V13      CIM-534      2572
V2      FH-2000      2273         V14      CIM-496      3531-i
V3      FH-1152      760-iv       V15      RH-512       2307
V4      NIAB-824     2633-vii     V16      NIAB-824     2554-xi
V5      MNH-732      2417         V17      ALSEEMI H    2779-iii
V6      MNH-700      2522         V18      CIM-476      2540
V7      MNH-768      2560         V19      FH-1000      2318
V8      RH-510       2393         V20      FH-2925      2360
V9      SLH-279      2671         V21      NIBGE-2      2425
V10     VH-148       2802-ii      V22      PB-843       2609
V11     BH-162       2478         V23      PB-899       2745
V12     BH-163       2339         V24      MJ-7         2569

Remained: i, iii, iv, vii at CRSS Khanewal, CRI, Fsd, CRSS Jhang,
CRS Sahiwal and PSC Khanewal, respectively.

Table 3: Yield performance (Kg/ha of NIAB-824 in DGR trials (2005-2006)

ode      Variety      Mean      Code      Variety      Mean

V4       FH-113       2358      V21       SLH-279      1988
V8       H-151        2336      V16       NIAB-884     1979
V11      MNH- 7862    210       V15       MNH-789      1969
V2       PB-899 21    29        V1        SLH-284      1967
V14      VH-148 21    26        V1        9N-4         1959
V6       SLH-317      2119      V3        NIAB-846     1951
V24      MJ-7         2045      V9        CIM-538      1859
V5       CIM-534      2041      V1        2BH-162      1846
V20      FH-901       2029      V10       FH-115       1845
V18      NIAB-824     2022      V23       RH-51        1840
V7       VH-156       2003      V22       MJ-6         1751
V13      FH-207       1998      V17       BH-164       1568

Table 4: Yield performance of NIAB-824 in NCVT trials (2005-2006)

Variety       Plant         Yield         Yield
              pop./ha       Kg/ha         Kg/ha

CRIS-466      39374         2809          3070
PB-899        40573         2864          3037
H-151-F2      40486         2845          3024
MNH-786       40220         2771          2964
CRIS-461      39948         2736          2947
TH-35/99      40721         2787          2945
NIAB-824      37233         2521          2913
CIM-534       40137         2689          2883
NIBGE-4       38235         2550          2870
MNH-789       39616         2634          2861

Variety       Plant         Yield         Yield
              pop./ha       Kg/ha         Kg/ha

FH-207        40394         2676          2850
BH-162        37483         2454          2817
CIM-538       39448         2553          2785
N-78/499      38807         2509          2782
NIAB-884      40205         2576          2757
GH-99         33737         2154          2747
MJ-7          40300         2542          2720
FH-115        38850         2435          2697
TH-84/99      38828         2096          2323

Calculated on standard plant population i.e. 43000/ha

Table 5: Field Response of Different Cotton Strains Against CLCuV in
DGR at NIAB, Faisalabad (2004-2005)

Strain %       Disease index    Disease Reaction

FH-2006        28.37            Highly Susceptible
FH-2000        10.07            Moderately susceptible
FH-115         20.48            Highly Susceptible
NIAB-884       12.86            Moderately susceptible
MNH-732        8.89             Moderately resistant
MNH-700        11.81            Moderately susceptible
MNH-768        11.21            Moderately susceptible
RH-510         3.92             Resistant
SLH-279        4.80             Resistant
VH-148         9.73             Moderately resistant
BH-162         12.31            Moderately susceptible
BH-163         16.28            Susceptible
CIM-534        28.06            Highly Susceptible
CIM-496        6.61             Moderately resistant
RH-512         8.76             Moderately resistant
NIAB-824       10.45            Moderately susceptible
ALSEEMI H      4.45             Resistant
CIM-476        8.66             Moderately resistant
FH-1000        20.67            Highly Susceptible
FH-2925        8.85             Moderately resistant
NIBGE-2        5.93             Moderately resistant
PB-843         5.0              Resistant
PB-899         15.33            Susceptible
MJ-7           8.70             Moderately resistant

Source: BSCV Project Rport

Table 6: Fiber quality characteristics of candidate variety
NIAB-884, NIAB-824 and NIAB-846

Variety       Length        Strength      Fineness      Maturity
              (mm)          (TPPSI)       (mg/inch)     (%)

NIAB-884      28.9          90.0          4.1           82.7
NIAB-824      27.0-30.0     94.0          4.0-4.6       83-90
NIAB-846      28.0-30.0     92.4          4.2-4.6       80.0-87.6

Table 7: Screening of candidate cotton varieties against
CLCuV through  grafting and under natural condition's in

Varieties     Through grafting          Under field conditions

              * Disease response        % Disease index

BT            H.S                       2.85
BH-162        H.S                       7.49
NIAB-884      H.S                       5.42
MJ-7          H.S                       5.21
TH-84/99      H.S                       21.87
FH-115        H.S                       14.67
NIAB-824      H.S                       4.01
MNH-789       H.S                       4.28
CIM-534       H.S                       12.42
FH-207        H.S                       8.04
CRIS-466      H.S                       10.65
MNH-786       H.S                       2.41
PB-899        H.S                       9.06
H-151-F2      H.S                       1.88
CRIS-461      H.S                       6.06
TH-35/99      H.S                       2.40
NIBGE-4       H.S                       7.25
CIM-499       H.S                       9.34
GH-99         H.S                       23.03
CIM-538       H.S                       16.93
SLH284        H.S                       26.70
PB-899        H.S                       11.60
NIAB846       H.S                       16.38
FH-113        H.S                       14.83
CIM-534       H.S                       13.74
SLH-317       H.S                       13.38
BH-156        H.S                       6.61
H-151         H.S                       19.92
CIM-538       H.S                       21.60
FH-115        H.S                       24.62
MNH-786       H.S                       3.41
BH-162        H.S                       18.14
FH-207        H.S                       8.72
VH-148        H.S                       18.82
MNH-789       H.S                       11.11
NIAB-884      H.S                       18.89
BH-164        H.S                       32.77
NIAB-824      H.S                       16.14
N-4           H.S                       19.67
FH-901        H.S                       9.38
SLH-279       H.S                       15.98
MJ-6          H.S                       13.43
RH-511        H.S                       13.69
MJ-7          H.S                       7.64
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Title Annotation:Original Articles
Author:Aslam, M.; Ashfaq, M.; Saeed, Tahir; Allah, Sami Ul; Sajjad, Muhammad
Publication:American-Eurasian Journal of Sustainable Agriculture
Article Type:Report
Geographic Code:9INDI
Date:Dec 1, 2009
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