Journal of Applied Botany and Food Quality 92, 7 - 14 (2019), DOI:10.5073/JABFQ.2019.092.002 1Department of Botany, Bharathiar University, Coimbatore, Tamil Nadu, India 2Department of Biotechnology, K.S. Rangasamy College of Technology, Tiruchengode, Namakkal District, Tamil Nadu, India 3Department of Biotechnology, VIT University, Vellore, Tamil Nadu, India Architectural effect of different tea clones on the development of blister blight disease Ponnusamy Ponmurugan1, Balasubramanian Mythili Gnanamangai2*, Kolandaisamy Manjukarunambika3 (Submitted: April 8, 2017; Accepted: February 20, 2018) * Corresponding author Summary An attempt has been made to analyze the architectural traits of six elite tea (Camellia sinensis) clones representing the three principal taxa Assam, China and Cambod with respect to the correlation of blister blight disease (Exobasidium vexans) development. In order to analyze the architecture, branching habit and flushing behavior were observed and subsequently compared with disease incidence. All the clones followed similar architectural pattern irrespective of the cultivar but varied with levels of disease severity. The number of branches was higher in China when compared to Assam and Cambod, branch length was bigger in Assam followed by Cambod and China. Branch angle of all the clones lay well within the described range of theoretical value of 45 to 90°. In general, internodal length was bigger in Assam followed by Cambod and China. These architectural characteristics determined the number of harvestable tea shoots in the bush canopy. China cultivars exhibited an erectophile type of leaf angle, which influenced effective net photosynthesis, transpiration rates and light penetration in leaves. These factors are playing important roles in a disease development strategy. This study should be useful for clonal selection for new clearings and re-planting areas. Moreover, plants breeding programmes for studying the yield and tea quality losses due to blister blight disease benefit from the findings herein. Keywords: Blister blight, Exobasidium vexans incidence, plant architecture, tea bush pattern. Introduction Tea plant architecture is the spatial distribution of different leaf composition covering scale leaf, scar of scale leaf, fish leaf, main- tenance foliage, mother leaf, 1st, 2nd and 3rd leaves and a bud, stems and flowers on that plant at a given time (Fig. 1). Tea (Camellia sinensis) is an evergreen plant that undergoes many cultural ope- rations like plucking, pruning, tipping and centering. Plucking of young harvestable shoots containing 2-3 leaves and a bud is being done and subjected to manufacture for tea powder. Young shoots are continuously harvested at 10-14 day intervals which affects the bush health significantly. Tea bushes are pruned to attack many pests and diseases (Barthakur, 2011). After 4-5 years, depending upon the nature of tea cultivars and climate, tea bushes undergo pruning which gives rise to new shoots. Those shoots are physiologically effective. Some other factors determining the bush health are altitude of the tea garden, climate, pruning and tipping heights, length of the pruning cycle and the system of plucking. Tea plant architecture depends on geometric, environmental factors and tea estate elevations influencing growth, including plucking patterns like manual plucking and shear harvesting and monsoon seasons. It is determined by an interaction between the intrinsic architecture defined by its genome and the characteristics of the population in which it grows (plant density). Tea plants are planted by single, double and triple hedge methods to get the maximum crop yield per hectare. These planting styles regulate the architecture of tea plant. Tea plant canopies are complex depending upon the style of planting (planting layout), tipping height and pruning types like light (80-90 cm from the ground level), medium (60-70 cm) and heavy pruning (15-25 cm). Several parameters can be used to characterize the tea canopy architecture which includes leaf area index, leaf distribution and number of harvestable shoots on plucking surface (Girardin et al., 1999). Tea being a monoculture crop is susceptible to a number of de- structive diseases and provides a stable microclimate for various diverse phytopathogens (Premkumar et al., 2008). Among the leaf diseases, blister blight disease caused by a fungus Exobasidium vexans Massee is one of the most damaging diseases of tea plants worldwide (Sinniah et al., 2016). It is an important foliar disease incapable of causing enormous crop loss throughout the tea growing regions of Asia, especially in India, Sri Lanka, Indonesia and Japan. E. vexans completes its life cycle in a short span of 11-28 days (Premkumar and BaBy, 2005). Thus, many generations of the fungus are completed during the favorable condition of the monsoon. The pathogen infects young succulent harvestable shoots leading to severe crop loss. The crop loss varies with the geographical locations and nature of tea clones and seedlings. It was estimated to be 33% in Sri Lanka (de Silva et al., 1992), 20-25% in Indonesia and as high as 35% in India (radhakriShnan and BaBy, 2004). In addition to crop loss the disease adversely affects the quality of made tea (PonmuruGan et al., 2016). Variation in several architectural traits combines to give species a distinctive visual appearance. But these different architectures were well correlated with different light- interception properties (FalSter and WeStoBy, 2003) rather its other metabolism and conduciveness to disease. Commercial tea population lies under three principal taxa, ‘China’ (Camellia sinensis (L.) O. Kuntze), ‘Assam’ (C. assamica ssp. assa- mica (Masters) Wight) and ‘Cambod’ (C. assamica ssp. lasiocalyx (Planch ex Watt) Wight) (Barua, 1965). Plant architecture is the visible, morphological expression of the genetic blue print of a tree (halle, 1978). Architectural analysis of tea clones will facilitate selection of the better traits against high yield, pests and disease resistance. The objective of the present study is to analyze the architectural features of different tea clones with respect to the development of blister blight disease over the course of the growing seasons. Materials and methods Six elite tea clones viz., UPASI-1 and UPASI-3 (Assam), UPASI-9 and UPASI-15 (China) and UPASI-17 and TRI-2025 (Cambod) maintained in UPASI Tea Research Institute Experimental Garden under similar cultural conditions were examined for various architectural parameters. The choice of the six tea cultivars was 8 P. Ponmurugan, B.M. Gnanamangai, K. Manjukarunambika selected on the basis of differences in architectural features, green leaf yield potential and tea quality parameters. Moreover, studies were undertaken to assess the specific effect of the architectural traits on epidemic blister blight development, tea cultivars with similar levels of susceptibility and resistance to blister blight were chosen. All the clones were planted in 1969 at 120 × 120cm spacing, in contour single hedge planting system. Observations on canopy architecture, floral characteristics and leaf morphology were made with unplucked tea plants maintained in the breeding plots and plants under regular cultural operations. Data on position of trunk, branch angle, branch length and number of branches were recorded. Branches were designated as the first order (n+1), the second order (n+2) and so on till seventh order (n+7). Internodal length was measured in the well-developed shoots which are about to be plucked. Shoot components were designated as a bud, first (7th order), second (6th order), third (5th order), mother (4th order), fish (3rd order), and scale (2nd order) leaves and maintenance foliage (1st order). Branch and internodal lengths were measured with a metric scale. Branch and leaf angles were measured with a circular ocular meter (honda and FiSher, 1978). Leaf area was also calculated conventionally. All the observations were replicated 10 times in separate bushes and the data were subjected to statistical analysis. Blister blight disease incidence was assessed every plucking round in all the three experimental blocks covering Assam, China and Cambod tea cultivars. Young tender shoots containing three leaves and a bud were collected randomly from the harvest baskets and each shoot was examined individually for the incidence of blister blight. Disease incidence was quantified on a percentage basis. The sequence of lesion development was divided into five stages such as 1) occurrence of translucent lesions, 2) well-defined lesions, 3) incipient stage of spore forming lesions, 4) vigorously sporulating lesions and 5) necrotized lesions and were carefully documented while examining the incidence of blister blight disease. Healthy and infected harvestable shoots were collected from all the bushes of six tea cultivars falling under Assam, China and Cambod traits periodically. The freshly harvested leaves were used for chlorophyll estimations. A 100 mg of harvested tea plant material was taken in preparing the leaf disc using the conventional paper puncher. The prepared leaf disc was submerged in 5 ml of DMSO (dimethyl sulfoxide) and the chlorophyll was estimated by following WellBurn (1994). For all other biochemical parameters the 1 gm of harvested shoots was extracted and concentrated with 80% ethanol, and the final extract was filtered and used for estimation of total sugars (AOAC, 1990), nitrogen (duBoiS, 1956), amino acids (moore and Stein, 1948), polyphenols (dev Choudhury and GoSWami, 1983) and catechins (SWain and hilliS, 1959). The data obtained were subjected to analysis of variance (ANOVA) and the significant means were segregated by critical difference (CD) at various levels of significance (Gomez and Gomez, 1984). Results Wide variations were observed in the canopy architecture among tea cultivars (Tab. 1). Assam tea cultivars were orthotrophic with ‘de- current’ or ‘deliquescent’ branches i.e., terminal leader eventually disappears and lateral branches tend to grow upwards. China tea cultivars were plagiotrophic with excurrent branches i.e., a prominent terminal leader with horizontal drooping lateral branches while Cambod tea cultivars were neither orthotrophic nor plagiotropic. All the clones bear identical branching dynamics with non-rhythmic or continuous growth without endogenous periodicity of extension. In China cultivars, the number of branches (second to fourth order) was significantly higher than in Assam and Cambod cultivars (Tab. 2). Significant variation in the branch length was observed between Assam and China tea cultivars while it was not significant either between Cambod and Assam or Cambod and China cultivars (Tab. 2). Branch angle lies well within the range of 45 to 90° (Tab. 2). Tea leaves follow a special pattern of arrangement, ‘anisophylly’. Each shoot comprises one or two scale leaves, one fish leaf, mother leaf and three to four normal leaves with a terminal bud (Fig. 1). No significant variation was observed between the shape and size of maintenance foliage, mother leaf and third leaf. Internodal length increased gradually from maintenance foliage to third leaf and de- clined in second and first leaves. Among the clones, no significant variation was observed in the intermodal lengths of maintenance Tab. 1: Comparison of architectural parameters of different tea cultivars Parameters Tea cultivars Assam China Cambod Habit Tree 10-15m tall Big shrub 1-3m tall Erect shrub 5-6m tall Growth habit Under trees with virgate branches Vigorous shrub with Erect, openly branched shrubs compound branches Branching pattern Semi-orthotrophic arise above the Semi-plagiotropic compact, Semi-orthotropic and semi- ground, excellent spread arise from the ground plagiotropic with profuse branching, arise from the base. No. of branches in each order 2.8 to 6.0 3.5 to 8.7 2.8 to 6.6 Branch length (cm) 11 to 36 7 to 25 8 to 30 Branch angle(°) 50 to 70 40 to 47 50 to 70 Internode length (cm) 2 to 6 1 to 4 1 to 5 Stem circumference (cm) 37.5 30.5 33.3 Length of main stem (cm) 37.5 28.8 32.5 Leaves Large, horizontal, broad, light green Small, narrow, erect, dark green medium size, broad, light green, semi-erect Leaf angle type Planophile (>70°) Erectophile (<50°) Oligophile (50-70°) Flowering Sparse Profuse Medium dense Plucking surface (cm2) 9788 9044 9358 Plucking points/sq.ft 155.5 125.5 141.0 Yield Potential High Low Moderate Tea quality Medium Superior Superior Blister blight due to leaf architechture 9 Bud Mother leaf Fish leaf Scale leaf Maintenance leaf Third leaf Second leaf First leaf Assam (9788 cm2, 155.5/sq.ft; respectively) followed by Cambod (9358 cm2 and 141/sq.ft) and China (9044 cm2 and 125.5/sq.ft). They may be attributed to leaf size and leaf distribution in the canopy. Assam cultivars had a higher leaf angle followed by Cambod and China (Tab. 3). Flowers are solitary, pleonanthic type where they are produced on lateral branches and thus the shoot are not limited by flowering. Flowering is always on old branches (rhamiflory) and so does not affect the shoot construction during its life cycle as observed in many species. Sympodial branching, sylleptic growth habit, semi- orthotropic and semi-plagiotropic branching, anisophylly, peonanthic flowers and rhamiflory are characteristic to Camellia spp. (Fig. 3B). Blister blight infection was more in Assam cultivar tea leaves than in Cambod and China, which is coinciding with the result of leaf angle and leaf area. Leaf angle of Assam cultivars was planophile (> 70°) type which were horizontal, whereas, the same in China and Cambod cultivars was erectophile (< 50°) and oligophile (50-70°) types; respectively. An average of leaf angle in China cultivars recorded was 28.1° in UPASI-9 and 27.7° in UPASI-15. Whereas, it was 54.2° and 56°; respectively, with Assam UPASI-1 and UPASI-3. The observations made in different tea cultivars with respect to blister blight disease incidence were presented in the Tab. 3. Dif- ferent stages of blister blight lesions were recorded in tea shoots. The lesions were fully matured and ready for spore dispersal in the third leaf (Fig. 3C) subsequently the lesion got necrotized (Fig. 3D). On the second leaf, however, the lesions were immature and slightly lesser in number. The first leaf showed fewer lesions, which were in their early stages of development. The pattern remained the same in all the three tea cultivars. Blister blight infection was registered the maximum with Assam UPASI-3 followed by Cambod TRI-2025 Tab. 2: Branching pattern parameters of various tea clones Branch parameters Assam China Cambod C.D. at P = 0.05 UPASI-1 UPASI-3 UPASI-9 UPASI-15 UPASI-17 TRI-2025 No. of Branches First order 5.7 5.5 6.7 6.5 6.6 4.5 1.3 Second order 6.3 6.7 8.7 8.3 6.4 5.7 2.1 Third order 4.1 3.7 6.8 6.5 4.6 3.5 1.3 Fourth order 4.2 4.6 7.2 5.4 4.6 3.4 1.5 Fifth order 2.7 3.0 3.9 3.4 3.5 2.8 1.1 Average 4.6 4.7 6.7 6.0 5.1 4.0 1.5 Branch length (cm) First order 23.8 22.7 19.9 24.0 28.4 20.2 6.3 Second order 20.8 23.7 29.4 32.5 34.4 24.5 7.5 Third order 31.0 28.7 22.4 28.5 25.2 24.2 6.5 Fourth order 10.7 12.5 19.3 20.5 16.4 14.6 6.2 Fifth order 08.5 07.5 10.5 15.3 08.4 08.8 3.3 Average 19.0 19.0 20.3 24.2 22.6 18.5 6.0 Branch angle(°) First order 67.3 53.5 44.5 43.5 52.3 65.7 18.3 Second order 70.5 57.8 44.3 43.5 62.7 70.7 25.2 Third order 72.8 63.6 43.6 45.0 53.0 55.5 17.2 Fourth order 51.5 53.5 42.5 40.0 52.5 60.7 13.5 Fifth order 63.5 58.5 42.5 45.6 50.7 55.5 16.5 Average 65.1 57.4 43.5 43.5 54.2 61.6 18.1 Fig. 1: Shoot structure of a periodic tea shoot containing different types of leaves leaf to scale leaf, scale leaf to fish leaf and fish leaf to mother leaf (Tab. 3). Tea canopies in terms of number of plucking points and plucking surface among tea cultivars vary at different levels. Both numbers of plucking points and plucking surface were found to be higher in Average 34.4 72.9 15.1 51.2 24.8 61.1 16.1 Figure 1. Shoot structure of a periodic tea shoot containing different types of leaves First order lateral Figure 2. Branching pattern of a tea bush Bud Mother leaf Fish leaf Scale leaf Maintenance leaf Third leaf Second leaf First leaf Third order lateral Second order lateral Internode Banji New growth 10 P. Ponmurugan, B.M. Gnanamangai, K. Manjukarunambika and minimum with China UPASI-15. These tea clones were highly susceptible to blister blight infection. In contrast, tea clones such as Assam UPASI-1, Cambod UPASI-17 and China UPASI-9 were tolerant to blister blight. Observations on the various stages of blister blight infections in tea leaves such as translucent, well- defined, spore forming, sporulating and necrotized lesions, there is no significant difference recorded (Tab. 3). Various degrees of biochemical constituents were observed in all the six tea cultivars (Fig. 4). Healthy and infected clone of the same cultivar showed significant reduction in all biochemical constituents, especially the chlorophyll, polyphenols and catechins. The difference was high between the healthy and infected clones of UPASI-15 followed by TRI-2025 and UPASI-3. In contrast the difference was comparatively low in all the biochemical parameters including sugar and nitrogen in UPASI-1, UPASI-9 and UPASI-17 clones. All the biochemical constituents in healthy conditions registered no significant difference among the six tea cultivars except for polyphenols which is high in UPASI -15. Discussion China cultivars having more branches indicate the vulnerability of the disease incidence. Pruning induced more number of proleptic and sylleptic branches with high shrubby appearance (zhou and hara, 1990) which lead to die back of weaker branches (PonmuruGan et al., 2000). Branch length was more in the case of second and third Tab. 3: Leaf architectural parameters of various tea clones and blister blight disease incidence Leaf parameters Assam China Cambod C.D. at P = 0.05 UPASI-1 UPASI-3 UPASI-9 UPASI-15 UPASI-17 TRI-2025 Leaf angle(0) First order 55.3 b 52.3 b 27.8 a 27.5 a 45.7 b 43.3 b 10.7 Second order 58.5 bc 60.5 bc 30.5 a 30.5 a 50.6 bc 43.5 b 11.6 Third order 55.7 bd 63.2 bd 31.5 a 28.8 a 42.5 bc 45.6 bc 10.6 Fourth order 72.5 bd 62.2 bd 27.0 a 25.7 a 45.2 bc 43.8 bc 13.3 Fifth order 56.5 bcd 65.5 bd 40.5 a 42.7 abc 55.6 bcd 60.5 bd 15.3 Sixth order 50.7 bc 52.6 bc 20.5 a 22.5 a 48.5 bc 47.3 bc 14.7 Seventh order 30.5 bcd 35.7 bd 18.7 a 16.0 a 23.5 abc 25.6 abcd 11.3 Average 54.2 56.0 28.1 27.7 44.5 44.2 12.5 Leaf area (cm2) Maintenance foliage (MF) 23.5ab 30.5c 22.2a 20.9a 20.5a 27.5bc 3.0 Scale leaf (SL) 00.8a 01.3a 00.7a 00.7a 01.3a 01.3a 1.1 Fish Leaf (FL) 00.7a 02.2b 01.3ab 01.1ab 01.5ab 01.5ab 1.1 Mother Leaf (ML) 19.6a 29.9d 19.9a 19.0a 19.5ac 23.0bc 2.5 IIIrd leaf (3L) 20.0b 29.0d 18.0a 19.0ab 18.0a 23.0c 1.6 IInd leaf (2L) 15.0bc 19.0d 13.5ab 11.1a 16.0b 17.5cd 2.5 Ist leaf (1L) 07.3bc 11.5be 06.8bc 04.9a 07.5bcd 08.3bd 1.3 Average 12.4 17.6 11.8 11.0 12.0 14.6 1.9 Internodal length (cm) MF><3L 4.9be 4.3bcde 3.7ac 3.0a 4.7bde 4.0bcd 0.7 3L><2L 7.5b 6.0bc 4.7ac 3.7a 6.0bc 5.5ac 2.2 2L><1L 4.5cd 4.7d 2.3ab 2.0a 3.3abc 3.5bcd 1.3 1L>