The effect of maternal environment on seed and seedling quality in tomato

Transcription

1 WAGENINGEN UR The effect of mternl environment on seed nd seedling qulity in tomto Agrwl, Vikrnt 6/25/2014 Supervised y: Wilco Ligterink, Shungchen Chen & Nfiseh Geshnizjni

2 Tle of contents Contents Tle of contents... 0 Astrct... 2 Introduction... 2 Mteril nd methods... 5 Plnt mteril... 5 Germintion ssy... 6 Seedling growth... 7 Dt nlysis... 7 Results... 8 Discussion Acknowledgement References... 40

3 Astrct Seed qulity is complex term which depends on genetic nd environmentl fctors. Growth nd development of plnt not only effected y existing environment condition. It lso depends on prentl environmentl condition such s light, temperture during seed development nd mturtion stge. We hve conducted fctoril experiment to know effect on seed nd seedling qulity in tomto on seed developed under different nitrte nd phosphte tretment. On seed developed under different nitrte nd phosphte condition of tomto RIL popultion nd prents, we hve mesured the germintion percentge, fresh root weight, dry root weight, fresh shoot weight nd dry shoot weight. We hve found tht effect of nitrogen nd phosphorous on seed qulity is genotype dependnt. For instnce in line 261 seed developed under very high nitrte showed less germintion on other hnd in line 221, 230 nd money showed higher germintion in seed developed under high nitrte condition. In line 254, 261, 290, money nd pimp no difference in different phosphte tretment with respect to fresh shoot weight. However fresh shoot weight found low in seed developed under very low phosphte condition in line 205, 221 nd 230 compre to other phosphte condition. Introduction Seed qulity is complex term which depends on genetic nd environmentl fctors[1]. During the seed developmentl stge, the seed qulity is governed y the genetic nd environment interctions [1, 2]. Seed germintion ility, dormncy, longevity, seedling vigour nd uniform growth re the importnt elements of seed qulity [1]. Severl definitions of seed qulity re given in literture, for instnce the cpility of seed to germinte in different environmentl conditions nd grow into helthy plntlets[3], seed qulity is the viility nd vigour elements of seeds which llows it to develop into helthy

4 seedlings under vrious environmentl conditions [4]. In prcticl terms, the definition of seed qulity lso depends on the type of industry. In food industry, seed qulity my depend on high strch nd oil content. In griculture, seed qulity depends on germintion nd seedling estlishment in vrious environmentl conditions [1]. Seed compnies focus on improving seed qulity t hrvest nd posthrvest stges y controlling hrvesting time nd posthrvest tretments such s sorting, grding nd priming etc. However, seed qulity cn lso e improved y controlling production environments[1]. As n exmple in studies on Aridopsis thlin, Elwell et l (2011) found tht progeny shows differences in seed weight, seed germintion nd erly seedling development stge when the sme genotype is grown in different environments [5][5]. Growth nd development of plnt not only effected y existing environment condition[5]. It lso depends on prentl environmentl condition such s light, temperture during seed development nd mturtion stge [6]. Vrious studies show the impct of production environments on seed qulity elements s descried elow. For the effect of temperture in Aridopsis thlin, Blonder et l. (2007) found tht seeds from prents grown t wrm temperture (25 C) showed quicker germintion nd root elongtion thn seeds from prents grown under cold conditions (15 C) [7]. Similr results were shown in study of P. Lnceolt [8]. In n experiment conducted y Demir et l (2004) in wtermelon (Citrullus lntus) dry mss nd longevity of seeds ws igger in cooler growing sites compred to wrmer growing sites. In nother experiment conducted y [9] with tomto (Solnum lycopersicum), seedling estlishment from seed hrvested in spring ws higher thn in utumn [10].

5 In tomto, long dy conditions during fruit ripening, results in seeds with less germintion thn seeds of mternl plnts exposed to shorter dy length [11]. Limited wter vilility during seed development hd little effect on seed qulity of Lctuc stiv L. Although, there ws n increse in seed weight ut there were fewer seed production per plnt [2]. On the other hnd, no significnt effect ws found on germintion nd primry seedling growth in tomto from seed developed under different irrigtion intervl [12]. For nutrient effect in Solnum lycopersicum, higher dose of nitrogen ppliction improve germintion of progeny [13]. Higher dose of nitrogen nd phosphorous in comintion to mother plnt cn help to improve the germintion rte nd seedling growth of progeny [14]. In experimentl study on Autilon theophrsti y Wulff et l 1992, they hve found tht the increse in mternl nutrient supply hs positive effect on erly stge of seedling growth of progeny [6]. However, the mternl nutrient effect lso depends on genotype nd progeny growth environmentl condition[6]. On other hnd, in the field experiment conducted y Singh nd Kumr 2010 on effect of phosphorus nd nitrogen fertiliztion on seed production of tomto, they hve found tht with increse mount of nitrogen ppliction showed positive effect on seed yield nd higher phosphorus ppliction show negtive effect on seed yield. However, no significnt effect of nitrogen nd phosphorus ws found with respect to the seed qulity [15]. The germintion percentge in Synpsis rvensis ws found to e decresed from seed progeny of prent grown under high nitrogen percentge [16]. High mount of nutrient nd dy light to mother plnt in Cmpnul mericn resulted in the high seed weight of the progenies ut low germintion percentge [17]. Seeds of Amrnthus retroflexus grown under high nitrogen level showed dormncy while tht on

6 low nitrogen level showed higher germintion rte. However, low nitrogen nd gierellic cid ppliction during seed development stge showed highest germintion percentge [18]. Different environmentl fctors ffect the seed nd seedling qulity in vrious crops s discussed ove. The im of this study is to exmine the effect of nitrogen nd phosphorous on seed nd seedling qulity in tomto. In erlier experimentl studies only one or two genotypes were studied. In this study, we used lines from recominnt inred line (RIL) popultion of Solnum lycopersicum cv. Moneymker nd Solnum pimpinellifolium nd the prents. Solnum lycopersicum is cultivted tomto species nd seed germintion nd erly seedling growth is sensitive to iotic stress. Solnum pimpinellifolium is wild tomto species with more tolernce to iotic stress during seed germintion nd erly seedling growth [19]. The use of this genetic vrition in our study llows us to not only see the effect of the mternl environment on seed nd seedling qulity, ut lso if there is genotype y environmentl interction for these trits. Mteril nd methods Plnt mteril Seed tches from the recominnt inred line (RIL) popultion of Solnum lycopersicum cv. Moneymker nd Solnum pimpinellifolium nd the prents re used in our experiments. The RIL lines were 205, 221, 230, 254, 261 nd 290. The RIL lines nd prents were grown under different phosphte nd nitrte concentrtion during flowering nd seed development (Tle 1).

7 Tle 1: The different nitrte nd phosphte concentrtions during growth of the RIL lines the prents of the RILs. Nitrte Phosphte Control 14mM 1000µM Very low nitrte 0.2mM 1000µM Low nitrte 3mM 1000µM High nitrte 20mM 1000µM Very high nitrte 36mM 1000µM Very low phosphte 14mM 30µM Low phosphte 14mM 100µM High phosphte 14mM 5000µM Very high phosphte 14mM 10000µM Germintion ssy The seeds were sown in trnsprent plstic trys (from DBP plstics, size 15 X 21cm) for germintion. Ech germintion try hd 2 lue filter ppers (Blue lotter pper, Anchor pper compny, size 5.6 X 8 inches) nd 50ml -0.5MP NCl. In ech try 3 smple of round 25 to 35 seeds were sown y using msk (Fig. 1). A rndomized complete lock design ws used nd 12 trys were piled up nd on the top nd the ottom of ech pile, one try contining 2 white filter ppers nd 50ml MQ wter were plced nd the top ws covered with lid to void unequl evportion. Then pile ws put in trnsprent plstic g nd plced t 4 C for 72 hours for strtifiction. Afterwrds the gs were trnsferred to n incutor t 25 C in light for germintion. The trys were kept in the incutor for 10 dys nd during the initil 3 dys pictures were tken of ech try twice dy. From the 4 th to 10 th dy pictures were tken once dy. Pictures were tken with Nikon D80 cmer

8 fixed to Kiser Stnd t 60cm distnce. The cmer ws connected to computer vi Nikon cmer control pro softwre version 2. Figure 1. The rrngement of three different seeds smples sown in germintion try Seedling growth Seedling growth ws mesured y the method descried y Khn et l (2012) [4]. In short, seedling growth ws mesured y performing germintion ssy s ove till time point 50 percent vile seed germintion on MQ insted of NCl. The first 10 germinted seeds were discrded per smple. From the remining germinted seeds 10 seeds per smple were trnsferred to Copenhgen tle t 22.5 C. The trnsferred seeds were llowed to grow for 10 dys fter which fresh weight of root nd shoot of ech smple. Susequently the shoots nd roots were dried in n oven t 80 C for 48 hours to otin dry root nd shoot weight. Dt nlysis Germintion trits were mesured y using the germintor pckge s descried in Joosen et. l 2010 [20] nd in more detiled mnul from the WSL. Different prmeters were otined. In this report the three prmeters used re t10 (time to germinte first 10% of vile seed), t50 (time to germinte 50% vile seed) nd gmax (mximum seed

9 gmax% germintion percentge) to nlyse the effect on seed germintion. In genotype if germintion percentge remin elow 10% tht genotype excluded from t10 sttisticl nlysis. Also in t50 sttisticl nlysis if germintion percentge remin elow 50% tht genotype excluded. To nlyse seedling trit s fresh nd dry shoot weight nd fresh nd dry root weight one wy ANOVA nlysis done in Genstt 16sp1 edition ws used. To check significnt effects of ech tretment fischers lest significnt difference test (LSD) ws used t =0.05. Results To see the effect of nitrte nd phosphte tretment during seed development on progeny germintion we hve mesured the progeny germintion percentge, t 50 nd t 10. The sme seeds were used to study the effect of nitrte nd phosphte tretment during seed development on progeny seedling growth. In this respect we hve mesured fresh root nd shoot weight nd dry root nd shoot weight of 10 dys old seedlings % 80% 70% 60% 50% 40% 30% 20% 10% 0% c c c c c c Tretment Figure 2 Germintion percentge (gmax%) nd SE ± in line 205 fter 10 dys incution t 25 C in -0.5MP slt concentrtion. Different letters indicte sttisticl significnt difference y student t test; α=0.05.

10 t10 In line 205 much lower germintion is found in the seeds of mother plnts treted with very high nitrte condition (15%) compre to seeds of control (76%), low nitrte (70%), high phosphte (78%) nd very high phosphte (76%) treted mother plnts. Among other tretments in reltion to control no significnt difference in germintion percentge ws oserved. However the germintion percentge ws more in seeds of high phosphte (78%) treted mother plnts thn seeds of very low nitrte (52%) nd high nitrte (54%) treted mother plnts (Fig.2). For line 205 we hve not oserved significnt differences etween different nitrte nd phosphte concentrtions on the t 50 (Dt not shown) c c c c c c 0.0 Tretment Figure 3. Time in hours nd SE ± for 10 percent vile seeds to germinte. In line 205 fter 10 dys incution t 25 C in -0.5MP slt concentrtion. Different letters indictes sttisticl significnt difference y student t test; α=0.05. For the t 10 we hve only oserved significnt difference etween seeds of very low phosphte treted mother plnt nd seeds of control nd high phosphte treted mother plnt. The seeds of very low phosphte treted mother plnt took hours, seeds of control hours nd seeds of high phosphte hours to rech t 10 (Fig.3).

11 gmax% 100% % 60% 40% 20% 0% c c c c c c Tretment Figure 4. Germintion percentge (gmax%) nd SE± in line 221 fter 10 dys incution t 25 C in -0.5MP slt concentrtion. Different letters indicte sttisticl significnt difference y student t test; α=0.05. In line 221 the seed germintion percentge in seeds of very high nitrte mother plnt (53%) ws higher thn seeds of very low (2%), low nitrte (4%) nd very low phosphte (26%) treted mother plnt. However with respect to control we hve not found ny significnt effect of phosphte nd nitrte on seed germintion percentge (Fig.4). For the t 10 we hve not oserved ny significnt differences in phosphte nd nitrte tretment of the mother plnt (Dt not shown). Vlues for t 50 were not clculted for line 221 due to too low germintion percentges.

12 gmax% gmax% % 30% 25% 20% 15% 10% 5% 0% Tretment Figure 5. The germintion percentge (gmax%) nd SE ± in line 230 fter 10 dys incution t 25 C in - 0.5MP slt concentrtion. Different letters indicte sttisticl significnt difference y student t test; α=0.05. In line 230 we hve oserved the germintion in seeds of very high nitrte treted plnt which ws 27% only. In other tretment we hve not oserved ny germintion (Fig. 5). Therefore, Line 230 is excluded from t 50 nd t 10 nlysis % 100% 80% 60% c c c c c 40% 20% 0% c Tretment Figure 6. The germintion percentge (gmax%) nd SE ± in line 254 fter 10 dys incution t 25 C in - 0.5MP slt concentrtion. Different letters indicte sttisticl significnt difference y student t test; α=0.05. In line 254 the 98% germintion in seeds of very high nd low nitrte treted mother plnt oserved. It ws higher compre to seeds of high nitrte nd low phosphte treted mother

13 t50 plnt. In seeds of high nitrte treted mother plnt 80% nd in seeds of low phosphte treted mother plnt 88% germintion oserved (Fig.6) d c c cd Tretment Figure 7. Time in hours nd SE ± for 50 percent vile seeds to germinte. In line 254 fter 10 dys incution t 25 C in -0.5MP slt concentrtion. Different letters indicte sttisticl significnt difference y student t test; α=0.05. For thet 50 nlysis we hve not oserved significnt differences with respect to control except seeds of high phosphte treted mother plnt. However we hve oserved the significnt differences for different phosphte nd nitrte tretment. Rech to t 50 seeds of high phosphte took 132 hours, very high nitrte took153 hours nd low nitrte took 152 hours, low phosphte took 168 hours nd high nitrte took 178 hours in order oserved (Fig.7).

14 gmax% t c d c cd Tretment Figure 8. Time in hours nd SE ± for 10 percent vile seeds to germinte. In line 254 fter 10 dys incution t 25 C in -0.5MP slt concentrtion. Different letters indicte sttisticl significnt difference y student t test; α=0.05. For the t 10 with respect to control (130.9hr) seeds of very high phosphte (122.4hr) treted mother plnt germinted erly. On other hnd seeds of low phosphte (137.2hr) treted mother plnt nd seeds of high nitrte (147hr) treted mother plnt germinted lter (Fig.8) % 100% 80% 60% 40% c c c 20% 0% Tretment Figure 9. The germintion percentge (gmax%) nd SE ± in line 261 fter 10 dys incution t 25 C in - 0.5MP slt concentrtion. Different letters indicte sttisticl significnt difference y student t test; α=0.05. In line 261except seeds of very low nitrte treted mother plnt with respect to control no significnt deference oserved. In line 261 the germintion ws mximum with seeds of

15 t10 mother plnt low nitrte treted (100%). However not significnt difference from seeds of low nitrte treted mother plnt nd seeds of high phosphte treted mother plnt (Fig.9). In line 261 in t 50 nlysis we hve not oserved ny significnt differences. Dt not shown c c c c c c 0.0 Tretment Figure 10 Time in hours nd SE ± for 10 percent vile seeds to germinte. In line 261 fter 10 dys incution t 25 C in -0.5MP slt concentrtion. Different letters indicte sttisticl significnt difference y student t test; α=0.05. For the t 10 nlysis with respect to control no significnt differences we hve oserved. However seeds of mother plnt with high phosphte treted (136hr) reched erly thn seeds of very low phosphte treted mother plnt (161hr), seeds of low nitrte treted mother plnt (164.7hr), seeds of very low nitrte treted mother plnt (164.3hr) nd seeds of high nitrte treted mother plnt (170.2hr) (Fig.10).

16 t50 gmax% % 80% 60% 40% 20% c c c c c c 0% Tretment Figure 11 The germintion percentge (gmax%) nd SE ± in line 290 fter 10 dys incution t 25 C in - 0.5MP slt concentrtion. Different letters indicte sttisticl significnt difference y student t test; α=0.05. In line 290 with respect to control (91%) nd seeds of very low phosphte treted mother plnt (82%) significntly less germintion oserved in seeds of very high phosphte treted mother plnt (56%), seeds of low (45%) nd very low nitrte treted mother plnt (47%). The effect of low phosphte, high nitrte nd very high nitrte not significntly different from others. (Fig.11) Tretment Figure 12. Time in hours nd SE ± for 50 percent vile seeds to germinte. In line 290 fter 10 dys incution t 25 C in -0.5MP slt concentrtion. Different letters indicte sttisticl significnt difference y student t test; α=0.05.

17 t10 For the t 50 nlysis the control (202.5hr) nd seeds of very low phosphte treted mother plnt (218.1hr) fster compre to seeds of high phosphte treted mother plnt (240.6hr) nd very high phosphte treted mother plnt (274.5hr) (Fig.12) c ce cde e d cde cd ce 0.0 Figure 13. Time in hours nd SE ± for 10 percent vile seeds to germinte. In line 290 fter 10 dys incution t 25 C in -0.5MP slt concentrtion. Different letters indicte sttisticl significnt difference y student t test; α=0.05. For the t 10 nlysis seeds of control (149.3hr) ws fster thn seeds of low nitrte (207.7hr), high nitrte (207.5hr) nd very high nitrte (230.6hr) treted mother plnt nd seeds of very high phosphte treted mother plnt (203.6hr). The seeds of very high nitrte treted mother plnt (230.6hr) delyed thn seeds of mother plnt treted with very low nitrte (149.3hr) nd very low phosphte treted mother plnt (177.3hr) (Fig.13).

18 gmax% gmax% 25% Moneymker 20% 15% 10% d d cd 5% 0% c c cd cd cd Tretment Figure 14 The germintion percentge (gmax%) nd SE ± in line money fter 10 dys incution t 25 C in - 0.5MP slt concentrtion. Different letters indicte sttisticl significnt difference y student t test; α=0.05. In money seeds of very high nitrte (17%) nd low phosphte (13%) treted mother plnt higher germintion percentge oserved thn seeds of very low nitrte (0%) nd low nitrte (0%) treted mother plnt. From t 50 nd t 10 nlysis money is excluded (Fig.14). Pimp 120% 100% 80% 60% 40% 20% c d cd d cd cd cd d 0% Tretment Figure 15 The germintion percentge (gmax%) nd SE ± in line pimp fter 10 dys incution t 25 C in - 0.5MP slt concentrtion. Different letters indicte sttisticl significnt difference y student t test; α=0.05.

19 t50 In line pimp the germintion percentge oserved low in seeds of very low nitrte treted mother plnt (47%) compre to seeds of very high nitrte treted mother plnt (98%) nd control (93%). No significnt different effect of other tretment oserved (Fig.17) Pimp Tretment Figure 16 Time in hours nd SE ± for 50 percent vile seeds to germinte. In line pimp fter 10 dys incution t 25 C in -0.5MP slt concentrtion. Different letters indicte sttisticl significnt difference y student t test; α=0.05 For the t 50 nlysis seeds of control (198.9hr) nd seeds of very high nitrte plnt (193.3) germinted fster thn seeds of high phosphte treted mother plnt (227.6hr). No significnt differences oserved for other tretment effect (Fig.16).

20 t Pimp d c cd cd c c c 0.0 Tretment Figure 17 Time in hours nd SE ± for 10 percent vile seeds to germinte. In line pimp fter 10 dys incution t 25 C in -0.5MP slt concentrtion. Different letters indicte sttisticl significnt difference y student t test; α=0.05 For the pimp in t 10 nlysis seeds of control (148hr) consumed less time thn seeds of low phosphte (167.6hr), high phosphte (184.5hr) nd very low phosphte (202.7hr) treted mother plnt nd seeds of very low nitrte (220.7hr) treted mother plnt. Seeds of very low nitrte (220.7hr) treted mother plnt required more time thn seeds of low nitrte (167.9hr) nd very high nitrte (153.1hr) treted mother plnt nd seeds of low phosphte (167.6hr), high phosphte (184.5hr) nd very high phosphte (174.3hr) treted mother plnt. The seeds of high phosphte (184.5hr.) treted mother plnt consumed more time thn seeds of very high nitrte treted mother plnt (174.3hr) (Fig.17). Result on seedling qulity trits fresh root nd shoot weight nd dry root nd shoot weight fter 10dys growth on Copenhgen tle in line 205, 221, 230, 254, 261, 290, money nd pimp.

21 Fresh shoot weight in grm c c c c c c 0 0.2_N 3_N 20_N 30_P 100_P 5000_P 10000_P Control Tretment Figure 18 The Mens nd SE± of fresh shoot weight in grm 10 dys fter growth on copenhgen tle in line 205. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05 In line 205 with respect to fresh shoot weight only in seeds of mother plnt treted under very low phosphte low shoot weight oserved compre to ll other nitrte nd phosphte tretment. Furthermore fresh shoot weight of seed of mother plnt treted under high nitrte oserved more thn seeds of mother plnt treted under very low nitrte condition (Fig 18).

22 Fresh root weight in grm Dry Shoot weight in grm _N 3_N 20_N 30_P 100_P 5000_P 10000_P Control Tretment Figure 19 The Mens nd SE± of dry shoot weight in grm 10 dys fter growth on copenhgen tle in line 205. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05. In line 205 for the dry shoot weight no significnt difference found in different tretment except seeds of very low phosphte treted mother plnt. The dry shoot weight from seeds of very low phosphte treted mother plnt found low compre to ll other phosphte nd nitrte tretment effects (Fig 19) c c c c c c 0.2_N 3_N 20_N 30_P 100_P 5000_P 10000_P Control Tretment Figure 20 The Mens nd SE± of fresh root weight in grm 10 dys fter growth on copenhgen tle in line 205. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05

23 Fresh shoot weight in grm In line 205 for the fresh root weight from seeds of very low phosphte treted mother plnt oserved low compre to seeds of low phosphte, high phosphte nd very high phosphte nd high nitrte nd control. The fresh root weight from seeds of high nitrte treted mother plnt found higher thn seeds of very low nitrte treted mother plnt (Fig 20). In line 205 with respect to dry root weight no significnt difference oserved. Dt not shown _N 20_N 36_N 30_P 100_P 5000_P 10000_P Control Tretment Figure 21 The Mens nd SE± of fresh shoot weight in grm 10 dys fter growth on copenhgen tle in line 221. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05. In line 221 for the fresh shoot weight no significnt difference found in different tretment except seeds of very low phosphte treted mother plnt. The fresh shoot weight from seeds of very low phosphte treted mother plnt found low compre to ll other phosphte nd nitrte tretment effects (Fig 21).

24 Dry shoot weight in grm _N 20_N 36_N 30_P 100_P 5000_P 10000_P Control Tretment Figure 22 The Mens nd SE± of dry shoot weight in grm 10 dys fter growth on copenhgen tle in line 221. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05 In line 221 for the dry shoot weight no significnt difference found in different tretment except seeds of very low phosphte treted mother plnt. The fresh shoot weight from seeds of very low phosphte treted mother plnt found low compre to ll other phosphte nd nitrte tretment effects (Fig 22). In line 221 with respect to fresh nd dry root weight no significnt difference oserved. Dt not shown.

25 Fresh shoot weight in grm c c c c c c Tretment Figure 23 The Mens nd SE± of fresh shoot weight in grm 10 dys fter growth on copenhgen tle in line 230. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05 In line 230 for the fresh shoot weight, in seeds of very low nitrte treted mother plnt less fresh shoot oserved compre to seeds of very high phosphte treted mother plnt. In seeds of very low nitrte, very low phosphte nd very high phosphte treted mother plnt less fresh shoot weight founded compre to seeds of low, high nd very high nitrte nd low nd high phosphte treted mother plnt nd control (Fig. 23).

26 Dry shoot weight in grm c c c c c Tretment Figure 24 The Mens nd SE± of dry shoot weight in grm 10 dys fter growth on copenhgen tle in line 230. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05. In line 230 for the dry shoot weight, low dry shoot weight oserved in seeds of very low nitrte treted mother plnt compre to seed of low nitrte nd high nitrte treted mother plnt nd low phosphte nd high phosphte treted mother plnt nd control. The dry shoot weight of seeds of very high nitrte treted mother plnt higher thn seeds of high nitrte nd very low phosphte treted mother plnt (Fig. 24).

27 Fresh root weight in grm cd c e c de Tretment Figure 25 The Mens nd SE± of fresh root weight in grm 10 dys fter growth on copenhgen tle in line 230. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05. In line 230 for the fresh root weight in seeds of very high nitrte treted mother plnt higher weight oserved thn seeds of very low nitrte, low nitrte nd high nitrte treted mother plnt nd very low phosphte, low phosphte, high phosphte nd very high phosphte treted mother plnt. In seeds of control nd low nitrte treted mother plnt higher weight oserved thn seeds of very low nitrte nd high nitrte treted mother plnt nd seeds of very low phosphte, low phosphte nd very high phosphte treted mother plnt. In seeds of very low nitrte treted mother plnt low weight oserved thn seeds of high nitrte, very low phosphte, low phosphte nd high phosphte treted mother plnt (Fig. 25).

28 Dry root weight in grm c cd c e c cde de Tretment Figure 26 The Mens nd SE± of dry root weight in grm 10 dys fter growth on copenhgen tle in line 205. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05. In line 230 for the dry root weight in seeds of very high nitrte treted mother plnt higher weight found thn very low nitrte, low nitrte nd high nitrte treted mother plnt nd in very low phosphte, high phosphte nd very high phosphte treted mother plnt. In seeds of low phosphte treted mother plnt nd control higher weight oserved thn seeds of high phosphte nd very high phosphte treted mother plnt. Higher dry root weight oserved in seeds of very high phosphte treted mother plnt thn seeds of high phosphte treted mother plnt. No significnt difference found in very low nitrte, low nitrte nd high nitrte nd very low phosphte nd very high phosphte tretment (Fig. 26). In line 254 with respect to fresh nd dry shoot weight nd fresh nd dry root weight no significnt effect of different tretment oserved. Dt not shown. In line 261 with respect to fresh shoot weight no significnt of different tretment oserved. Dt not shown.

29 Dry shoot weight in grm c cd cd cd c c d 0 Tretmetn Figure 27 The Mens nd SE± of dry shoot weight in grm 10 dys fter growth on copenhgen tle in line 261. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05 In line 261 for the dry shoot weight in seeds of control higher dry shoot weight oserved thn seeds of low nd very low nitrte treted mother plnt nd seeds of very low phosphte, high phosphte nd very high phosphte treted mother plnt. In seeds of low nitrte, high nitrte nd very high nitrte nd low phosphte, high phosphte nd very high phosphte higher dry shoot weight oserved thn seeds of very low nitrte nd seeds of very low phosphte treted mother plnt. In seeds of very low phosphte treted mother plnt lower dry shoot weight found thn seeds of very low nitrte treted mother plnt (Fig 27). In line 261 with respect to fresh root weight no significnt of different tretment oserved. Dt not shown.

30 Dry root weight in grm c c c c c Tretment Figure 28 The Mens nd SE± of dry root weight in grm 10 dys fter growth on copenhgen tle in line 261. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05. In line 261 for the dry root weight in seeds of high nitrte treted mother plnt higher dry root weight oserved thn seeds of very low nitrte nd low nitrte treted mother plnt nd very low nd high phosphte treted mother plnt. In seeds of high nitrte, low phosphte nd very high phosphte nd control higher dry root weight oserved thn seeds of high phosphte treted mother plnt (Fig 28). In line 290 with respect to fresh shoot weight no significnt effect of different tretment oserved. Dt not shown.

31 Dry shoot weight in grm d c c c c cd c Figure 29 The Mens nd SE± of dry shoot weight in grm 10 dys fter growth on copenhgen tle in line 290. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05 In line 290 for the dry shoot weight higher dry root weight oserved in seeds of high nitrte treted mother plnt thn very low nitrte, low nitrte nd very high nitrte treted mother plnt nd very low phosphte, low phosphte nd high phosphte treted mother plnt nd control. Lower dry shoot weight oserved in seeds very low nitrte treted mother plnt thn seeds of control, very high nitrte treted mother plnt nd very low phosphte, low phosphte, high phosphte nd very high phosphte treted mother plnt (Fig 29). In line 290 with respect to fresh nd dry root weight no significnt effect of different phosphte nd nitrte tretment oserved. Dt not shown.

32 Dry shoot weight in grm Fresh shoot weight in grm Money Tretment Figure 30 The Mens nd SE± of fresh shoot weight in grm 10 dys fter growth on copenhgen tle in line money. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05 In line money for the fresh shoot weight lower fresh shoot weight found in seeds of very low nitrte nd very high nitrte treted mother plnt thn seeds of control, low nitrte nd high nitrte nd low phosphte, high phosphte nd very high phosphte treted mother plnt (Fig 30) Money c c c c c c c 0 Tretment Figure 31 The Mens nd SE± of dry shoot weight in grm 10 dys fter growth on copenhgen tle in line money. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05

33 Fresh root weight in grm In line money for the dry shoot weight, lowest dry shoot weight oserved in seeds of very low nitrte treted mother plnt compre other phosphte nd nitrte tretment effect. In seeds of very high nitrte treted mother plnt dry shoot weight higher thn seeds of very low phosphte treted mother plnt (Fig 31). Money Tretment Figure 32 The Mens nd SE± of fresh root weight in grm 10 dys fter growth on copenhgen tle in line money. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05. In line money for the fresh root weight only in seeds of very high nitrte treted mother plnt higher weight oserved thn other nitrte nd phosphte tretment effect (Fig 32). In line money with respect to dry root weight nd no significnt effect of different tretment oserved. Dt not shown. In line pimp with respect to fresh nd dry shoot weight no significnt effect of different tretment oserved. Dt not shown.

34 Fresh root weight in grm Pimp cd d d c cd d d Tretment Figure 33 The Mens nd SE± of fresh root weight in grm 10 dys fter growth on copenhgen tle in line pimp. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05. In line Pimp for the fresh root weight lowest fresh root weight found in seeds of very high phosphte treted mother plnt compre to other phosphte nd nitrte tretment. In seed of control, high nitrte nd high phosphte treted mother plnt higher fresh root weight oserved thn seed of seed of very high phosphte nd very low phosphte treted mother plnt (Fig 33). In line pimp with respect to dry root weight nd no significnt effect of different tretment oserved. Dt not shown.

35 Fresh shoot weight in grm e d cd c c c Money Pimp Control Figure 34 The Mens nd SE± of fresh shoot weight in grm 10 dys fter growth on copenhgen tle in ll genotype under control condition. Different letters indicte sttisticlly significnt differences y Fisher s lest significnt difference (LSD) test; α=0.05. This is the effect from ll line s mother plnt grown in control condition. Fresh shoot weight highest in line money nd lowest in line 221. This indictes the genotypic vrition while seed developed under sme control tretment (Fig 34). 0.2N 3N 20N 30P 100P 5000P 10000P Control 3N N P P P P Control N Figure 35. Person s correltion of different nitrte nd phosphte tretments sed on fresh shoot weight. Colour represent the degree of correltion, green colour represent very high correltion, yellow represent high correltion, ornge represent low or negligile correltion nd red represents negtive correltion.. The very low nitrte tretment hd minute correltion with tretment high nitrte, low phosphte, high phosphte nd control. Wek positive correltion with tretment very high nitrte nd very high phosphte nd wek negtive reltionship with very low phosphte. The very low phosphte hd high positive correltion with ll other tretment except very

36 high nitrte nd very low nitrte. The very high nitrte hd moderte correltion with ll the tretments except very low nitrte nd very low phosphte tretment. The remining tretment low nitrte, high nitrte, low phosphte, high phosphte, very high phosphte nd control hd very strong high positive correltion with ech other (Fig. 35). 0.2N 3N 20N 30P 100P 5000P 10000P Control 3N N P P P P Control N Figure 36 Person s correltion of different nitrte nd phosphte tretment sed on fresh root weight. Colour represent the degree of correltion, green colour represent very high correltion, yellow represent high correltion, ornge represent low or negligile correltion nd red represents negtive correltion. The tretment low phosphte hd moderte positive correltion with high nitrte nd very high phosphte nd high correltion with very low nitrte tretment. It hd low negtive correltion with control nd minute positive correltion with high phosphte nd very high nitrte. The very low nitrte tretment hd strong high positive correltion with high nitrte nd very high phosphte, very low with control nd high positive correltion with low nitrte, low phosphte, high phosphte nd very high phosphte. the low nitrte tretment hd very high positive strong correltion with high phosphte, very high phosphte, control nd very high nitrte, high positive correltion with high nitrte nd very low phosphte nd minute positive correltion with low phosphte tretment. The high nitrte hd high very high strong positive correltion with very low phosphte nd high positive correltion with high phosphte nd very high phosphte nd low correltion with control nd low phosphte tretment. The tretment very low phosphte hd high strong correltion with

37 very high phosphte nd very high nitrte, high correltion with low nd high phosphte nd low correltion with control. The high phosphte hd very strong high correltion with very high nitrte nd high correltion with very high phosphte nd control tretment very high phosphte hd moderte correltion with control (Fig. 36). Discussion In previous study to know the effect of nitrte nd phosphte on seed qulity in tomto experiment conducted only using one genotype. In this study we hve found the vrition in seed qulity prmeters sed on genotypes nd tretment. For germintion percentge the effects were not the sme in ll genotypes. For nitrte tretment in line 261 seed developed under very high nitrte showed less germintion. In contrst, line 221, 230 nd money showed higher germintion in seeds developed under high nitrte conditions. For phosphte tretment we hve found higher germintion in seeds developed under very low phosphte conditions in line 290 nd higher germintion in seeds developed under low phosphte. However no significnt effect of different phosphte tretments on seed germintion ws found in line 205, 221, 230, 254, 261 nd money. On other hnd in previous study y Singh nd Kumr on tomto cultivr Ark Ah they did not find significnt effect of nitrogen nd phosphorous tretment on seed germintion [15]. The reson my e they hve only used the one cultivr. In our study we hve found tht higher germintion in seed developed under medium phosphte nd high nitrte condition. In nother study on tomto cultivr Moneymker showed tht the germintion rte nd seedling emergence ws higher in seeds developed under high nitrte nd high phosphte conditions [14]. However the experiment ws conducted on field condition. For t 10 nd t 50 we found the contrdictory result in line 254 nd 290. In line 254 seed developed in very

38 high phosphte condition require less time to rech, ut in 290 seed developed in very low phosphte condition require less time. For seedling qulity lso we hve found the different effect due to genotype nd tretment interction. For fresh shoot weight in line 221, 254, 261, 290 nd pimp no difference in different nitrte tretment ws mesured. However low fresh shoot weight ws mesured in line 205, 230 nd money from seed developed under very low nitrte condition. In line 254, 261, 290, money nd pimp no difference in different phosphte tretment ws mesured. However in line 205, 221 nd 230 low fresh shoot weight mesured from seed developed under very low phosphte condition compre to low phosphte, high phosphte nd very high phosphte condition. For dry shoot weight In line 205, 221, 254 nd pimp no difference in different nitrte tretment ws mesured. However low dry shoot weight ws mesured in line 230, 261, 290 nd money from seed developed under low nitrte condition compre to low nitrte, high nitrte nd very high nitrte condition. In line 230, 254, 290, money nd pimp no difference in different phosphte tretment ws mesured. However In line 205, 221 nd 261 lower dry shoot weight ws mesured from seed developed under low phosphte condition thn seed developed under low phosphte, high phosphte nd very high phosphte condition. For fresh root weight in line 221, 254, 261 nd 290 no significnt effect of different nitrte tretment ws mesured. However higher fresh root weight oserved in line 205, 230, money nd pimp from seed developed under very high nitrte condition compre to other very low nitrte, low nitrte nd high nitrte condition. In line 221, 230, 254, 261, 290 nd money no significnt effect of different phosphte tretment ws mesured. However in line 205 lower fresh root weight mesured from seed developed under very low phosphte

39 condition, ut in line pimp lower fresh root weight mesured from seed developed under high phosphte nd low phosphte condition. For dry root weight in line 205, 221, 254, 290, money nd pimp no significnt effect of different nitrte tretment ws. However in line 230 higher dry root weight oserved in seed developed under very high nitrte condition, ut in line 261 higher dry root oserved in seed developed under high nitrte condition. In line 205, 221, 254, 290, money nd pimp no significnt effect of different phosphte tretment ws mesured. However in in line 230 nd 261 low dry root weight oserved in seed developed under high phosphte condition. On tomto very few study hs een done so fr on the effect of seed developed under different nutrient conditions on seed qulity. In other crops some studies hve shown effect nd some not. In n experiment with two verities of Sorghum icolor L., significnt effect of seed development under different nitrte nd phosphte conditions on seed germintion hs een found, ut only in one vriety significnt effect on seedling emergence ws found. The germintion ws higher in seeds developed under comintion of norml nitrte nd phosphte conditions thn without nitrte nd phosphte [21]. In study on six cultivrs of rpeseed the uthor showed the significnt effect on germintion in seed developed under different nitrogen condition in field experiment. The higher germintion in cultivr zfrn in high nitrogen condition. However in lortory conditions significnt effect on germintion ws not found [22]. In study on Pisum stivum L., cultivr sprite uthor conducted two field experiment, in one experiment the higher seed yield ws mesured from plnt developed under high nitrte nd phosphte condition ut in susequent experiment no significnt effect on seed yield ws mesured, ut in oth experiment higher dry weight in seed developed under high nitrte nd phosphte tretment oserved [23]. However experiment conducted on only one vriety. In experiment on three cultivr of cotton (Gossypium

40 hirsutum L.), higher seed cotton yield in plnt developed under high nitrogen condition ws mesured thn plnt developed in low nitrogen condition [24]. In n experiment on chickpe (Cicer rientinum L.) using six cultivr, no significnt effect on 1000 grin weight in seed developed under different nitrogen nd phosphorous tretment ws mesured [25]. In field experiment on Cpsicum nnuum vr. nnuum L. cultivr Anheim Chili, uthor not found significnt effect of different nitrte tretment on seed qulity prmeter : germintion percentge, seedling weight nd field emergence [26]. However the experiment ws conducted only on one vriety. In field experiment on three vriety of okr uthor did not find ny significnt effect of nitrogen nd phosphorous on progeny seed germintion [27] There is n effect on mternl tretment of different phosphte nd nitrte condition. However we hve not find in ll genotype. Although we found the differences due to different phosphte nd nitrte tretment on seed qulity, it ws genotype dependnt. In this study we hve only used less repliction. In future for further study to revel more inside we need to increse repliction so we cn increse sttisticl power. This knowledge is giving indiction tht it is possile to identified gens which interct with nitrte nd phosphte to improve seed qulity. Acknowledgement My sincere thnks to Wilco Ligterink, Shungchen Chen, Nfiseh Geshnizjni, Elise Serin, Leo Willems, Jurin Rienstr nd Henk Hilhorst, who mde this work possile.

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