VEGETOS: An International Journal of Plant ResearchOnline ISSN: 2229-4473
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Research Article, Vegetos Vol: 30 Issue: 3

Towards Development of Hybrid Tossa Jute (Corchorus olitorius L.) using Chemical Hybridizing Agent: Insight into AgroMorphological and Reproductive Response

Sharma HK*, Kumar AA, Choudhary SB, Satya P, Maruthi RT and Karmakar PG

Crop Improvement Division, ICAR-Central Research Institute for Jute and Allied Fibres, Barrack pore, Kolkata- 700120, West Bengal, India

*Corresponding Author : Hariom Kumar Sharma
Crop Improvement Division, ICAR-Central Research Institute for Jute and Allied Fibres, Barrackpore, Kolkata- 700120, West Bengal, India
E-mail: [email protected]

Received: October 27, 2017 Accepted: May 31, 2017 Published: June 03, 2017

Citation: Sharma HK, Kumar AA, Choudhary SB, Satya P, Maruthi RT, et al. (2017) Towards Development of Hybrid Tossa Jute (Corchorus olitorius L.) using Chemical Hybridizing Agent: Insight into Agro-Morphological and Reproductive Response. Vegetos 30:3. doi: 10.5958/2229-4473.2017.00151.3

Abstract

Three male gametocides (maleic hydrazide, benzotriazole, surf excel detergent) were tested on tossa jute cv. JRO-204 for induction of male sterility in randomized block design with three replications. Effect of different treatments was assessed on different plant parts including morphological traits (seed yield and attributing traits) and reproductive parts (anther and stigma). Phytotoxic effects associated with different male gametocides were observed on different vegetative and reproductive parts. Visual toxic effects included stunted growth, leaf burning, delayed flower initiation and deformity of flower buds. Amongst different treatment maleic hydrazide induce more sterility with less phytotoxic effects on jute plant..

Keywords: Corchorus olitorius, Chemical hybridizing agents, Male sterilit

Introduction

Tossa jute (Corchorus olitorius L.) the second important fibre crop after cotton cultivated in West Bengal, Bihar, Odisha, Assam, Tripura, Meghalaya, Uttar Pradesh, Andhra Pradesh, Tamil Nadu and Maharashtra states of India [1]. India is the largest producer of jute followed by Bangladesh. Till 2009 fifteen varieties of tossa jute and sixteen varieties of white jute have been released for commercial cultivation in India [2] largely by exploiting additive genetic variance through pedigree selection. Although ~50% better parent heterosis for fibre yield have been reported in the crop [3] but remained untapped. As a result jute productivity found stagnated around 30-35 q/ha during last decade. The phenomenon coupled with sky shooting agrochemical input price seriously compromising economic viability of the crop and led to nationwide shrinking of jute acreage. In 2013-14 total 11083 thousand bales of jute fibre was produced from 756 thousand hectares area in India [4]. Despite of periodical upward revision of minimum support price of jute by government jute failed to attract farmers even in the traditional areas like West Bengal and Bihar. Improvement of national productivity of jute has been found only lasting solution for the problem. In this endeavor exploring prospects of utilizing untapped potential of heterosis by developing hybrids in the crop will prove well timed technological intervention. Since jute has hermaphrodite flowers. Therefore, regulation of pollination is prerequisite for the much talked hybrid development in the crop. With the non-availability of male sterile line in jute plant genetic resources and practical difficulties in in-vitro regeneration and genetic transformation of jute [5] applications of chemical hybridizing agents (CHA) would be an effective alternative due to their proven efficiency as male gametocides/ male sterilant/ pollen suppressant [6]. Many CHA (Maleic hydrazide, FW-450, ethrel, GA3, 2,4-D, NAA, DPX- 377, arsenicals and patented compounds, detergents) were reported to induce male sterility [6-11] General physiological and histo-chemical changes in response to these gametocides are well documented in cotton, sunflower, mustard and sesame by various workers. Chauhan and Kinoshita [7] studied the histo-chemical changes associated with treatment of male gametocides (MH and FW450) and reported deficiency of DNA, proteins and carbohydrates in tapetum layer of anthers of male sterile plants which resulted in production of nonviability of pollen grains in Sesamum indicum. Tripathi and Singh [10] observed degenerated and dysfunctional cell organelles in tapetum layer of sterile anthers of sunflower. Singh and Chouhan [12] reported suppression or overexpression of anther fertility specific genes (Bcpl and callase) based on RAPD, Dot blot and RTPCR techniques in Brassica juncea. From application point of view, among all thee CHA maleic hydrazide found the most commonly used male gametocides which have been reported to induce nearly complete sterility with less phototoxic effects [7,8]. Benzotriazole and Surf Excel detergent have also been used in Helianthus annuus, Brassica juncea, Raphanus sativus, Capsicum annuum and Gossypium arboreum for induction of male sterility [8.10,11,13]. However, available literature in jute is silent about effects of CHA in growth and reproductive fitness of jute that seriously compromising fate of hybrid development in the crop. To bridge the gap we studied effect and efficiency of three CHA chemicals namely maleic hydrazide (MH), benzotriazole (BTZ) and Surf Excel® (SE) detergent on agro-morphological and reproductive attributes of tossa jute (cv. JRO 204).

Materials and Methods

Tossa jute cv. JRO 204 used for induction of male sterility in the present study. The experiment was laid out in randomized block design (RBD) with three replications of 31 treatments (Table 1) in experimental field of ICAR-Central Research Institute for Jute and Allied Fibres (ICAR-CRIJAF), Barrackpore, Kolkata, India. Sowing was done in last week of August. Row to row spacing was 50 cm while plant to plant spacing was maintained at 5-7cm after thinning the plants at 30 days after sowing (DAS). Standard package of practices were followed to raise the crop. Initially, three chemicals namely maleic hydrazide (50, 100, 200, 300, 400, 500 ppm), benzotriazole (0.1, 0.3, 0.5, 0.7, 1.0, 2.0 %), and Surf excel® (0.5, 1.5, 2.0, 4.0 %) detergent powder (Hindustan Unilever Ltd.) were assayed for induction of male sterility. Since, higher doses of maleic hydrazide (300, 400, 500 ppm) produced deformed flower buds, and of benzotriazole (2.0%) as well as Surf excel ® (2%, 4%) caused stunted growth led by burning of meristametic tissues. Therefore, these doses of corresponding chemicals excluded from further study. As a result three doses of maleic hydrazide (50, 100, 200 ppm), five doses of benzotriazole (0.1, 0.3, 0.5, 0.7, 1.0%) and two doses of SE (0.5, 1.5%) were used in subsequent study that constituted treatments. Each treatment sprayed over ten plants in morning. For application of different treatments three growth stages were selected [S1: before flower initiation (35DAS), S2: after flower initiation, S1S2: spray at both stages]. Appropriate precautions were taken during application of treatments to prevent spray of chemicals on plants of adjacent treatments. Control was sprayed with distilled water. All treatments were applied using small hand sprayer [13].

  Chemical-concentration-stage-no. of sprays Days to flowering Plant height (cm) Number of branches Dry pod wt./plant (g) Pods/ plant Pod length (cm) Seeds/ pod Seed yield/ plant (g) 1000 seed wt (g)
1 MH-50-S1-3 54bcde 165.07ij 10def 16.2defg 42.36klmn 54.7ef 182.13fgh 7.60g 1.33b
2 MH-50-S2-2 54bcdef 159.58ij 10.53ef 20.78g 37.00hijklm 57.9def 199.6g 7.93g 1.43b
3 MH-50-S12-4 53bcd 157.33hij 8.19abcdef 15.2defg 37.67ijklm 56.1cdef 182.47fgh 7.07fg 1.23b
4 MH-100-S1-3 55bcdef 135.73efghi 8.13abcdef 16.4defg 33efghijklm 54.5cdef 181.37fgh 6.00efg 1.25b
5 MH-100-S2-2 58bcdefgh 157.53hij 6.8abcdef 18.13efg 33.1efghijklm 52.5bcdef 195.27gh 7.00fg 1.41b
6 MH-100-S12-4 54bcdef 158.92hij 10def 14.73cdefg 35.6ghijklm 51.7bcdef 152.23defgh 5.22bcdefg 1.05ab
7 MH-200-S1-3 60defghi 133.67efhi 5.6abcde 13.87bcdefg 23.4bcdefghi 54.5cdef 165.17efgh 5.33cdefg 1.24b
8 MH-200-S2-2 59cdefghi 147.53ghij 4.8abc 15.6defg 30.07defghijklm 54.2cdef 121.73cde 5.73defg 1.26b
9 MH-200-S12-4 61fghi 114.75defg 5.27abcd 15.13defg 33.87efghijklm 49.6abcdef 147.57defgh 4.72bcdefg 1.07ab
10 BTZ-0.1-S1-2 53bcd 153.97hij 9.97def 19.75fg 39.07jklm 57.0cdef 165.87efgh 6.17fg 1.37b
11 BTZ-0.1-S2-1 55bcdefg 161.88ij 8.82bcdef 19.4fg 45.17mn 57.7cdef 165.87efgh 6.55fg 1.34b
12 BTZ-0.1-S12-2 53bcd 136.87efghi 10.67ef 17.93efg 38.1jklm 58.9cdef 175.23efgh 6.87fg 1.18ab
13 BTZ-0.3-S1-2 53bcd 122.17defgh 9.8cdef 15.47defg 17.4abcd 54.5cdef 164.2efgh 6.53fg 1.14ab
14 BTZ-0.3-S2-1 53bc 101.63cde 10.68 18.8fg 44.13lmn 60.6cdef 123.57cde 6.50fg 1.32b
15 BTZ-0.3-S12-2 55bcdefg 93.8bcd 9.55cdef 15.35defg 36.4ghijklm 59.4cdef 146.73defgh 4.63abcdefg 1.15ab
16 BTZ-0.5-S1-2 58bcdefgh 105.8cdef 9.19cde 10.43abcd 20.02abcde 56.9cdef 132.47cdef 4.17abcdefg 1.12ab
17 BTZ-0.5-S2-1 55bcdefg 134.75efghi 7.33abcdef 13.3bcdefg 21.83abcdefg 51.5bcdef 146.53defgh 5.50defg 1.33b
18 BTZ-0.5-S12-2 54bcde 116.2defg 6.42abcdef 10.68abcdef 18.28abcd 54.6cdef 104.87bcd 3.92abcdefg 1.10ab
19 BTZ-0.7-S1-1 52ab 68.72abc 7.52abcdef 8.22abcde 22.47abcdefgh 50.6abc 89.8abc 3.03abcdef 1.07ab
20 BTZ-0.7-S2-1 52b 46.72a 5.23abcd 7.33abcd 15.84abcd 43.4abcde 84.1abc 1.78abcde 1.09ab
21 BTZ-1.0-S1-1 55bcdefg 48.83a 7.08abcdef 2.83a 20.33abcdef 36.5bcdef 51.07a 1.18abc 1.03ab
22 BTZ-1.0-S2-1 54bcd 49.33a 7.33abcdef 5.16abc 14.83abc 37.5abcd 54.67ab 1.56abcd 1.07ab
23 BTZ-1.5-S1-1 62hi 37.89a 3.22a 2.78a 8.05a 33.3ab 36.33a 0.44acdefg 0.99ab
24 BTZ-1.5-S2-1 61eghi 58.47ab 6.87abcdef 4.71ab 14.00ab 37.3ab 83.97abc 1.00ab 0.55a
25 SE-0.5-S1-2 65i 142.17fghij 3.85abdef 14.43bcdefg 29.28cdefghijk 55.2a 164.7efgh 6.17fg 1.33b
26 SE-0.5-S2-1 62ghi 155.82hij 5.08abcd 16.33defg 34.87fghijklm 55.7ab 177.83fgh 6.87fg 1.38b
27 SE-0.5-S12-2 61fghi 150.72ghij 5.2abcd 16.98efg 37.34ijklm 55.9cdef 160.46efgh 5.68defg 1.22b
28 SE-1.5-S1-2 61fghi 138.05efghi 3.65a 14.07bcdefg 28.53bcdefghijk 58.1cdef 147.23defgh 5.12bcdefg 1.22b
29 SE-1.5-S2-1 54bcdef 143.9ghij 5.33abcdef 13.22bcdefg 24.93bcdefghij 52.8cdef 150.1defgh 5.60defg 1.32b
30 SE-1.5-S12-2 57bcdefgh 140.76fghij 4.73abc 12.95bcdefg 35.58ghijklm 60.2cdef 143.73defg 5.09bcdefg 1.16ab
31 Control 45a 175.6j 10.53ef 22.4g 54.33n 61.4f 183.03fgh 7.73g 1.6b

Table 1: Effect of different CHA on mean performance of tossa jute variety JRO-204.

For testing pollen grains viability, unopened floral buds were collected from ten plants of each treatment and transferred into standard fixative [CarnoyA: alcohol and acetic acid solution (3:1 v/v)] followed by transfer into alcohol solution (70%) after 24 h. Finally anthers were removed from buds and pressed gently with the help of forceps and needle to ooze out pollen grains on slide. Observations were taken under light microscope after staining pollen grains with acetocarmine solution (0.5%). Minimum three microscopic fields were photographed for each treatment and further used for counting of sterile (unstained) and fertile (red stained) pollen grains. For testing of female sterility flower buds were cross pollinated with pollen grains from control plants after application of treatments. After pollination crossed flowers were collected in standard fixative after 1h of pollination. After 24h Collected samples were treated with 8 M NaOH for 1 h, and stained with 0 .1 % Aniline Blue dissolved in 0.1 N K3P04 for 2 h and viewed under fluorescent microscope. ANOVA for different traits was performed using SPSS version 18 software (SPSS Inc., Chicago, USA) and all graphs were prepared in Microsoft Excel 2010.

Results and Discussion

Impressive impact of heterosis on crop productivity improvement have inspired researcher to explore feasibility of developing hybrids in jute. Given the absence of genetic male sterile source and recalcitrant nature of the crop, application of chemical hybridizing agent to induce male sterility seems the most viable alternative in the endeavor. In this backdrop present investigation was designed to ascertain gametocidal potentiality of three chemicals namely MH, BTZ and SE in jute. Post application morphological phytotoxic effects of these CHA at different concentration recorded and analysed in one of the leading tossa jute cultivar namely JRO 204. Visual toxic effects included stunted growth, leaf burning, delayed flower initiation and deformity of flower buds (Figure 1A-1D). These phytotoxic effects are similar to the findings of Guan [14] and Liu [15] which reported abnormal development of reproductive organs and other plant parts. ANOVA revealed significant differences among different treatments (Table 2) indicating that different treatments significantly affected performance of different traits under study which is very well depicted with wide range of performance of traits. Highest CV (%) was recorded for seed yield/plant (24.71%) and branches/plant (21.63%) indicating that these two traits were highly affected by different treatments. Lowest CV (4.66%) was observed for days to 50% flowering indicating the trait is least affected by different doses of gametocides. Effect of different treatments on seed yield and attributing traits is depicted in (Figure 1A-1D). Effects of BTZ and SE was inversely proportional to the concentration of these chemicals as higher doses of BTZ were found to be highly toxic and led to reduced plant height, branches, seeds/ pod, pods/plant, pod length and poor seed yield/plant. Compared to control, application of CHA resulted into delayed flowering of treated plants irrespective of chemical nature. In control plant 50% flowering was attained on 45 DAS while in treated plant it was extended upto 65DAS (BTZ0.5_S1-2). Effect of higher doses of BTZ is clearly visible from stunted growth of plant (Figure 2). The finding supplements observation of Chouhan et al. (2005) reported reduction in number of fruits/plant, fruit size, total yield and delayed flowering with higher concentration of BTZ in chili, radish and cotton.

Traits MS Mean  ± SE(m) Range CV (%)
Days to 50% flowering 52.08** 55.90 ± 1.84 46.50-60.50 4.66
Plant height (cm) 4924.54** 123.01 ± 6.32 37.89-175.60 8.90
No. of branches 16.46** 7.34 ± 0.91 3.20-10.70 21.63
Dry pod weight/ plant (g) 83.41** 13.69 ± 1.56 2.80-22.40 19.83
No. of pods/ plant 350.31** 29.90 ± 2.43 8.06-54.33 14.12
Pod length (mm) 175.68** 51.9 ± 3.02 33.30-61.40 10.09
No. of seeds/pod 5703.73** 141.29 ± 9.70 36.33-199.60 11.89
Seed yield/ plant (g) 12.98** 5.11 ± 0.73 0.44-7.93 24.71
1000-seed weight (g) 0.103** 1.20 ± 0.11 0.55-1.60 16.27

Table 2: Effect of gametocides treatments on different quantitative traits of tossa jute.

Figure 1: Pollen sterility induced by different CHA treatments on tossa jute plant.

Figure 2: Morphological response of tossa jute plant to CHA. Treatment number indicates different doses of CHA (1= MH-50, BTZ-0.1, SE-0.5; 2= MH-100, BTZ-0.3, SE-1.5; 3= MH-200, BTZ-0.5; 4=BTZ-0.7; 5=BTZ-1.0; 6=BTZ-1.5)

Differential response on pollen viability was observed for different gametocides. Almost all three gametocides induced nearly complete sterility when they were applied on S12 stage (Figure 3). Higher doses of MH (200ppm) and SE (1.5%) not only induced male sterility but also caused floral deformity (Figure 1C-1D). However, such deformity was temporal as after some period normal flower buds were observed. Similar male sterility induction efficiency of SE (1.0, 2.0, 3.0%) were reported by Tripathi and Singh (2008b) in Helianthus annuus. To assess the effect of CHA on gynoecium viable pollen grains from control plants were used to pollinate the CHA treated plants after 24h of treatment. In order to test the effect of CHA on pistil of jute plant florescent microscopy of CHA treated plant was done after following artificial cross pollination using fertile pollens from control plant. Normal pollen germination was observed on MH100 treated plant (Figure 4) indicating stigma receptivity. Amongst all the treatments MH100 was found to be the safest as it induced 92.67-99.7% male sterility (Figure 1E, 3) when applied on S1 (92.67 %) and S12 (96.48%) stages without any yield compromise as seed yield of treated plants were found to be at par with control plant. Thus, MH100 has minimum possible phytotoxicity on jute growth and development except microsporogenesis. The finding together with observations of Singh et al. (1989) in cotton suggests that MH can be an ideal gametocide for future jute hybrid development programme.

Figure 3: Vegetative and reproductive effects of CHA application on tossa jute plant (A) burning symptoms on plant due to surf excel® @2%, 4% (B) stunted growth of benzotriazole treated plants @ 2% (C-D) deformed flower buds after CHA treatments (E) sterile pollen grains of maleic hydrazide treated plants.

Figure 4: Pollen germination response on pistil of MH100 treated tossa jute plant after artificial pollination.

Conflict of Interest

Authors declare no competing interest in manuscript.

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