A Morphometric and Geo Morphometric Study Comparing Two Sub Species of Taphrina Caerulescens and Taphrina Deformans

Oak leaf blister caused by Taphrina caerulescens (Figure 1&2) is endemic to North America. In 2015 in a study of the phytosanitary status of the Sierra Fría of Aguascalientes symptoms very similar to those that are caused by Taphrina were observed on numerous oak species by Moreno Taphrina diseases are best known in Europe and North America. Taphrina deformansthe most notorious species of this genus produces the deformation of leaves and fruits and later the defoliation of the


Introduction
), which results in the production of smaller fruits and fruit fall. When the disease is severe, it can result in a loss of 50% or more of the fruits. The disease can also affect the buds and twigs of plum and peach, resulting in the weakening of these fruit trees. The disease is more severe in the southeastern Gulf of the United States of America Sinclair et al. [1].
T. caerulescens is closely related to Taphrina deformans, which causes blisters on leaves and peach fruits, the asca of these two pathogens are indistinguishable, however, T. deformans infects species of peach trees whereas T. caerulescens only infects the Oaks The most important economic losses, however, are those produced by Taphrina deformans in peach, almond, nectarine and sometimes on plum. Many investigators claim that there is no difference between the ascus formed by T. caerulescens in oak species and T. deformans which affect prune and other stone fruits.
Little work has been done to confirm the similarly of differences that exist within the species and between the different species of Taphrina. In this study we propose the following objectives I.
Difference between the asci of T. caerulescens and T. deformans II. Difference between the asci of T. caerulescens taken from leaves of Quercus potosina and Quercus eduardii III. And finally compare the three groups

Selection of samples
Samples of asci of Taphrina caerulescens were isolated from leaves of Q potosina and Q eduardii and were selected at random from prepared semi-permanent and permanent slides of leaves infected with the disease. The sample photographs of Taphrina deformans Asci ( Figure 4) were downloaded from trusted websites; the photos downloaded were also downloaded randomly.   Photographs of ascus ( Figure 5&6) were edited using the Photoshop software package and were photographed using a Nikon D3000 which was mounted on a compound microscope (Leica DMS) at a magnification of 400x. After images were processed the configurations of landmark coordinates were scaled, translated and rotated by a generalized Procrustes analysis (GPA) using MorphoJ, TPSutil, TPS relw and TPS dig programs Klingenberg (4) & Rohlf [5]. A total of 107 landmarks were used per image.

Data analysis and statistic
Multivariate Analysis of Variance (MANOVA) Rohlf [5] was used to explore individual elemental fingerprint differences between the different isolates analyzed, followed by the Paired hoteling test to further test for differences between the isolates collect from the two oak species. Previously, normality and homogeneity of variance were tested (Shapiro-Wilk test, p > 0.05 and Shapiro-Wilk test, p > 0.05, respectively).
This geometric analysis was performed using 107 landmarks, reconstructed from distance measurements among Agricultural Research & Technology: Open Access Journal the landmarks. Shape variables generated from the landmark analysis were considered to be invariant regarding mathematical differences in translation, rotation, and scale Márquez et al. [6]. The multivariate regression of shape; size was computed as centroid size (CS), the square root of the sum of squared distances from each landmark to the specimen's centroid Loy et al. [7]. The relative warps (RW) were used to construct a matrix and a PCA was performed (relative warp analysis, RWA) in order to describe major trends in shape variations Márquez et al. [6] & Zelditch et al. [8].    The results for the images obtained from the TPs subjected to Procrustes fittings (Figure 7-9) shows that although all images follow the same pattern, there is notable more deformation in the Taphrina deformans image when compared to the other samples to the average image ( Figure 10). This visual analysis is confirmed for after a MANOVA was done for the three samples (p > 0.05) Q eduardii: Q potosina (0.091958); p < 0.05 Q eduardii: P persica (0.0005858); (p < 0.05) 0.0005858 P. persicae. eduardii, (p < 0.05) 3.75E-05 P. persicae: Q. potosina. Results are displayed in Table 1.  The difference between the samples is also seen when the main principal components were plotted on a graph (Table 2) The Paired Hoteling however showed a slight difference between the samples taken from the two oak species (T 2 :35.786); (F:2.4283); (p :0.046105) see results in Table 3.

Discussion
Results the geometric morphometric indices showed high separation between the ascus of T. deformans and T. caerulescens. This is very visible and very obvious in the graph where the main principal components are plotted. However, the results also show some degree of separation between the two isolates of T caerulescens, these results are in keeping with the already published works of other investigators principally Mix in hoes monograph of the Genera Taphrina. It is however important to determine the analytical tools to be used in these types of experiment.
when the PCR scores were analyzed using MANOVA it only showed that the T caerulescens isolates differed significantly when compared to T deformans, this test however did not show differences between the T. caerulescens isolates, however on comparing the T. caerulescens isolates with the Paired Hoteling (p :0.046105) it did show some level of separation.
This means that it would be possible perform monitoring studies without the need to apply the multiple techniques to show separation, to show differences among isolates of the same species the paired Hoteling method can be used alone. And to identify differences among various species a MANOPVA analysis of the PCR scores can be successfully used. This can help reduce the duplication of work, effort and possibly the cost in some cases; this is in keeping with observations made by Aviglianoa et al. [9].
In conclusion, this study is the first part in refuting other studies which claim that there are no differences between the ascus of T. deformans and T. caerulescens it also partially supports the claims that although there is some lever of separation, among the isolates of T. caerulescens they are still relatively similar in terms of their shapes. More study is recommended however to compare more isolates of Taphrina caerulescens on other host oak species of the Sierra Fria, Aguascalientes [10].